LOCUS       sequence-442617-        5679 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pAAV (non-flex) for expressing OCaMP, an orange calcium indicator, 
            in mammalian cells and in vivo. Contains hSyn promoter and RSET 
            leader sequence peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5679)
  TITLE     Sensitive orange calcium indicators for imaging neural activity
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5679)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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            "Unpublished"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-NES-OCaMP-WPRE
FEATURES             Location/Qualifiers
     source          1..5679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     protein_bind    137..157
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        179..626
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    626..650
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             723..740
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             744..776
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(780..788,789..803)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             819..875
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1571..1590
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2058..2082)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2092..2680
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2145..2165)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2563..2574)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2710..2727)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2716..2923
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2931..3071
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2931..3060
                     /label=AAV2 ITR
     rep_origin      3146..3601
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3233..3252)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3443..3464
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3618..3637)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3737..3759
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3797..3815)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3883..3987
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3988..4056,4057..4848)
                     /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(4206..4225)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5019..5607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5508..5527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5669..5679,1..130)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tctagacaac tttgtataga aaagttgctg cagagggccc tgcgtatgag
      181 tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc
      241 ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga
      301 gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga
      361 cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc
      421 tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg
      481 ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg
      541 accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag
      601 gagtcgtgtc gtgcctgaga gcgcagcaag tttgtacaaa aaagcaggct gccaccatgc
      661 tgcagaacga gcttgctctt aagttggctg gacttgatat taacaagact ggaggaggtt
      721 ctcatcatca tcatcatcat ggtatggcta gcatgactgg tggacagcaa atgggtcggg
      781 atctgtacga cgatgacgat aaggatcgat ggggatcccg tcgtaagtgg aacaaggcag
      841 gtcacgcagt cagagctata ggtcggctga gctcacccgt ggtttccgag cggatgtacc
      901 ccgaggacgg cgtcctgaag agcgagatca agaaggggct gaggctgaag gacggcggcc
      961 actacgccgc cgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     1021 cctacatcgt cgacataaag ttggacatcg tgtcccacaa cgaggactac accatcgtgg
     1081 aacagtgcga acgcgccgag ggccgccacc ccaccggcgg cagggacgag ctgtacaagg
     1141 gaggtacagg agggagtctg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
     1201 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     1261 agggcgaggg cgagggccgc ccctacgagg cctttcagac cgctaagctg aaggtgacca
     1321 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcaca tacggctcca
     1381 aggcctacat taagcaccca gccgacatcc ccgactactt taagctgtcc ttccccgagg
     1441 gcttcaggtg ggagcgcgtg atgaacttcg aggacggcgg cattattcac gttaaccagg
     1501 actcctccct gcaggacggc gtattcatct acaaggtgaa gctgcgcggc accaacttcc
     1561 cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggctacg cgtgaccaac
     1621 tgactgaaga gcagatcgca gaatttaaag aggctttctc cctatttgac aaggacgggg
     1681 atgggacgat aacaaccaag gagctgggga cggtgttccg gtctctgggg cagaacccca
     1741 cagaagcaga gctgcaggac atgatcaatg aagtagatgc cgacggtgac ggcacattcg
     1801 acttccctga gttcctgacg atgatggcaa gaaagatgaa tgacacagac agtgaagaag
     1861 aaattagaga agcgttccgc gtgtttgata aggacggcaa tggctacatc ggcgcagcag
     1921 agcttcgcca cgtgatgaca gaccttggag agaagttaac agatgaggag gttgatgaaa
     1981 tgatcagggt agcagacatc gatggggatg gtcaggtaaa ctacgaagag tttgtacaaa
     2041 tgatgacagc gaagtaaacc cagctttctt gtacaaagtg ggaattccga taatcaacct
     2101 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2161 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2221 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2281 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2341 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2401 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2461 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     2521 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2581 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     2641 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgggaatt cctagagctc
     2701 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2761 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2821 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2881 gcaaggggga ggattgggaa gagaatagca ggcatgctgg ggagggccgc aggaacccct
     2941 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     3001 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     3061 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3121 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3181 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     3241 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3301 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3361 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3421 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     3481 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     3541 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     3601 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     3661 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3721 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3781 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3841 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3901 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3961 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4021 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4081 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4141 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4201 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4261 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4321 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4381 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4441 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4501 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4561 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4621 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4681 aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt
     4741 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4801 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4861 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4921 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4981 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5041 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5101 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5161 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5221 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5281 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5341 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5401 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5461 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5521 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5581 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5641 gccttttgct ggccttttgc tcacatgtcc tgcaggcag
//
