LOCUS       sequence-492026-        6683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6683)
  AUTHORS   Minderer M, Liu W, Sumanovski LT, Kugler S, Helmchen F, Margolis DJ
  TITLE     Chronic imaging of cortical sensory map dynamics using a genetically
            encoded calcium indicator.
  JOURNAL   J Physiol. 2011 Nov 14.
  PUBMED    22083602
REFERENCE   2  (bases 1 to 6683)
  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: "J Physiol. 
            2011 Nov 14."
COMMENT     Sequence Label: AAV-6P-SEW-YC3.6
FEATURES             Location/Qualifiers
     source          1..6683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      474..483
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(480..482,483..485,486..998)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIE"
     CDS             1620..1697
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1704..2438
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
                     HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     misc_feature    2461..3049
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2514..2534)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2932..2943)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3082..3099)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3088..3295
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3304..3433
                     /label=AAV2 ITR
     rep_origin      3541..3996
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3628..3647)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3838..3859
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4063..4082)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4182..4204
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4242..4260)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4328..4432
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4433..4501,4502..5293)
                     /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(4651..4670)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5464..6052
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5953..5972
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6495..6543
                     /note="synthetic polyadenylation signal"
     misc_feature    6557..6648
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6597..6616
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gaattcaagc tgctagcgca agtgccacca
      481 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      541 gcgacgtaaa cggccacagg ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
      601 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
      661 tcgtgaccac cctgacctgg ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
      721 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgt accatcttct
      781 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
      841 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
      901 agctggagta caactacatc agccacaacg tctatatcac cgccgacaag cagaagaacg
      961 gcatcaaggc ccacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     1021 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1081 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     1141 tgctggagtt cgtgaccgcc gcccgcatgc atgaccaact gacagaagag cagattgcag
     1201 agttcaaaga agccttctca ttattcgaca aggatgggga cggcaccatc accacaaagg
     1261 aacttggcac cgttatgagg tcgcttggac aaaacccaac ggaagcagaa ttgcaggata
     1321 tgatcaatga agtcgatgct gatggcaatg gaacgattta ctttcctgaa tttcttacta
     1381 tgatggctag aaaaatgaag gacacagaca gcgaagagga aatccgagaa gcattccgtg
     1441 tttttgacaa ggatgggaac ggctacatca gcgctgctca gttacgtcac gtcatgacaa
     1501 acctcgggga gaagttaaca gatgaagaag ttgatgaaat gataagggaa gcagatatcg
     1561 atggtgatgg ccaagtaaac tatgaagagt ttgtacaaat gatgacagca aaggggggga
     1621 agaggcgctg gaagaaaaac ttcattgccg tcagcgctgc caaccggttc aagaagatct
     1681 ccagctccgg ggcactggag ctcatggacg gcggcgtgca gctcgccgac cactaccagc
     1741 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     1801 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1861 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggtggc agcggtggca
     1921 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1981 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2041 gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2101 tcgtgaccac cctgggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc
     2161 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2221 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2281 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2341 agctggagta caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg
     2401 gcatcaaggc caacttcaag atccgccaca acatcgagta agaggcgcaa gcttatcgat
     2461 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2521 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2581 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     2641 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     2701 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     2761 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2821 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     2881 gcctgtgttg ccacctggat tctgcgcggg acgcccttct gctacgtccc ttcggccctc
     2941 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3001 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt
     3061 cgactagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     3121 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3181 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3241 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggagagat
     3301 ctgaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3361 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     3421 agcgagcgcg cagagaggga gtggcccccc cccccccccc cccctgcagc ctggcgtaat
     3481 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     3541 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3601 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3661 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3721 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3781 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3841 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3901 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3961 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     4021 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     4081 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     4141 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     4201 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     4261 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     4321 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4381 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4441 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4501 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4561 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4621 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4681 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4741 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4801 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4861 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4921 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4981 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5041 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5101 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5161 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5221 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5281 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5341 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5401 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5461 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5521 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5581 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5641 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5701 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5761 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5821 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5881 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5941 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6001 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6061 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6121 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6181 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     6241 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcaggc tgcagggggg
     6301 gggggggggg gggggccact ccctctctgc gcgctcgctc gctcactgag gccgcccggg
     6361 caaagcccgg gcggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
     6421 cactaggggt tcctcagatc taggatcacg cgttctagaa atattaaggt acgggaggta
     6481 cttggagcgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     6541 gtgaatcgat agtactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag
     6601 caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctctat tctagactcg
     6661 agctgcagag ggccctgcgt atg
//
