LOCUS       sequence-474733-        5452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of iCre and mRuby3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5452)
  TITLE     Dendritic compartment specific spine formation in layer 5 neurons 
            underlies cortical circuit maturation during adolescence
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5452)
  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            "Unpublished"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-hsyn1-iCre-P2A-mRuby3
FEATURES             Location/Qualifiers
     source          1..5452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   38..167
                     /label=AAV2 ITR
     promoter        249..696
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      705..714
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             717..737
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1773..1829
                     /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             1830..2537
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTDI
                     ALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAVA
                     KYSNLGGGMDELYK"
     polyA_signal    2564..2685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2575..2594)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2629..2648
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2847..2976
                     /label=AAV2 ITR
     rep_origin      complement(3213..3801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3293..3312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3972..4763,4764..4832))
                     /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     4595..4614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4833..4937)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4963..5418)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5100..5121)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5312..5331
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agggcgatcg gtgcgggcct cttcgctatt acgccagctg cgcgctcgct cgctcactga
       61 ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga
      121 gcgagcgcgc agagagggag tggccaactc catcactagg ggttccttgt agttaatgat
      181 taacccgcca tgctacttat ctacgtagcc atgctctagg aagatctctg cagagggccc
      241 tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg
      301 acgaccgacc ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc
      361 ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca
      421 gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact
      481 gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt
      541 cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg
      601 cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg
      661 ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga attcgccacc atggtgccca
      721 agaagaagag gaaagtctcc aacctgctga ctgtgcacca aaacctgcct gccctccctg
      781 tggatgccac ctctgatgaa gtcaggaaga acctgatgga catgttcagg gacaggcagg
      841 ccttctctga acacacctgg aagatgctcc tgtctgtgtg cagatcctgg gctgcctggt
      901 gcaagctgaa caacaggaaa tggttccctg ctgaacctga ggatgtgagg gactacctcc
      961 tgtacctgca agccagaggc ctggctgtga agaccatcca acagcacctg ggccagctca
     1021 acatgctgca caggagatct ggcctgcctc gcccttctga ctccaatgct gtgtccctgg
     1081 tgatgaggag aatcagaaag gagaatgtgg atgctgggga gagagccaag caggccctgg
     1141 cctttgaacg cactgacttt gaccaagtca gatccctgat ggagaactct gacagatgcc
     1201 aggacatcag gaacctggcc ttcctgggca ttgcctacaa caccctgctg cgcattgccg
     1261 aaattgccag aatcagagtg aaggacatct cccgcaccga tggtgggaga atgctgatcc
     1321 acattggcag gaccaagacc ctggtgtcca cagctggtgt ggagaaggcc ctgtccctgg
     1381 gggttaccaa gctggtggag agatggatct ctgtgtctgg tgtggctgat gaccccaaca
     1441 actacctgtt ctgccgggtc agaaagaatg gtgtggctgc cccttctgcc acctcccaac
     1501 tgtccacccg ggccctggaa gggatctttg aggccaccca ccgcctgatc tatggtgcca
     1561 aggatgactc tgggcagaga tacctggcct ggtctggcca ctctgccaga gtgggtgctg
     1621 ccagggacat ggccagggct ggtgtgtcca tccctgaaat catgcaggct ggtggctgga
     1681 ccaatgtgaa cattgtgatg aactacatca gaaacctgga ctctgagact ggggccatgg
     1741 tgaggctgct cgaggatggg gacggatccg gagccacgaa cttctctctg ttaaagcaag
     1801 caggagacgt ggaagaaaac cccggtcctg tgtctaaggg cgaagagctg atcaaggaaa
     1861 atatgcgtat gaaggtggtc atggaaggtt cggtcaacgg ccaccaattc aaatgcacag
     1921 gtgaaggaga aggcagaccg tacgagggag tgcaaaccat gaggatcaaa gtcatcgagg
     1981 gaggacccct gccatttgcc tttgacattc ttgccacgtc gttcatgtat ggcagccgta
     2041 cctttatcaa gtacccggcc gacatccctg atttctttaa acagtccttt cctgagggtt
     2101 ttacttggga aagagttacg agatacgaag atggtggagt cgtcaccgtc acgcaggaca
     2161 ccagccttga ggatggcgag ctcgtctaca acgtcaaggt cagaggggta aactttccct
     2221 ccaatggtcc cgtgatgcag aagaagacca agggttggga gcctaataca gagatgatgt
     2281 atccagcaga tggtggtctg agaggataca ctgacatcgc actgaaagtt gatggtggtg
     2341 gccatctgca ctgcaacttc gtgacaactt acaggtcaaa aaagaccgtc gggaacatca
     2401 agatgcccgg tgtccatgcc gttgatcacc gcctggaaag gatcgaggag agtgacaatg
     2461 aaacctacgt agtgcaaaga gaagtggcag ttgccaaata cagcaacctt ggtggtggca
     2521 tggacgagct gtacaagtaa gcggccgctt cgagcagaca tgataagata cattgatgag
     2581 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     2641 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     2701 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac
     2761 ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg tagataagta
     2821 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc
     2881 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     2941 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc attaatgaat cggccaacgc
     3001 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3061 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3121 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3181 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3241 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3301 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3361 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3421 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3481 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3541 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3601 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3661 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3721 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3781 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3841 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3901 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3961 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4021 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4081 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4141 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4201 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4261 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4321 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4381 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4441 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4501 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4561 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4621 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4681 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4741 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4801 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4861 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4921 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     4981 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     5041 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     5101 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     5161 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     5221 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     5281 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     5341 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     5401 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg ga
//
