LOCUS       sequence-259370-        6474 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV expression of ultrasensitive protein calcium sensor from CamKII 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6474)
  TITLE     Penn Vector Core Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6474)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pENN.AAV.CamKII.GCaMP6f.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..6474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        149..1437
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1462..1471
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1468..1476,1477..1494,1495..1497,1498..1530,1531..1533,
                     1534..1557,1558..1584,1585..1641,1642..1647,1648..1917,
                     1918..1935,1936..2370,2371..2376,2377..2820)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      1564..1573
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1854..1875
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1981..2002)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2242..2261)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2833..3421
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2886..2906)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3304..3315)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3464..3585
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3475..3494)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3529..3548
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3737..3877
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3737..3866
                     /label=AAV2 ITR
     rep_origin      3952..4407
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4039..4058)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4249..4270
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4424..4443)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4543..4565
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4603..4621)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4689..4793
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4794..4862,4863..5654)
                     /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(5012..5031)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5825..6413
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6314..6333
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttaaca ttatggcctt aggtcacttc atctccatgg
      181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc
      241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa
      301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag
      361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca
      421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt
      481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc
      541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca
      601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt
      661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg
      721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag
      781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt
      841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca
      901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg
      961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt
     1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg
     1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag
     1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa
     1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa
     1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc
     1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt
     1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctct
     1441 agagcggtac cggatccacc cgccaccatg ggttctcatc atcatcatca tcatggtatg
     1501 gctagcatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga cgataaggat
     1561 ctcgccacca tggtcgactc atcacgtcgt aagtggaata agacaggtca cgcagtcaga
     1621 gctataggtc ggctgagctc actcgagaac gtctatatca aggccgacaa gcagaagaac
     1681 ggcatcaagg cgaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     1741 taccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1801 tacctgagcg tgcagtccaa actttcgaaa gaccccaacg agaagcgcga tcacatggtc
     1861 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc
     1921 ggtaccggag ggagcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1981 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggtgag
     2041 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     2101 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     2161 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacatccag
     2221 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     2281 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     2341 aacatcctgg ggcacaagct ggagtacaac ctgccggacc aactgactga agagcagatc
     2401 gcagaattta aagaggaatt ctccctattt gacaaggacg gggatgggac aataacaacc
     2461 aaggagctgg ggacggtgat gcggtctctg gggcagaacc ccacagaagc agagctgcag
     2521 gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc tgagttcctg
     2581 acaatgatgg caagaaaaat gaaatacagg gacacggaag aagaaattag agaagcgttc
     2641 ggtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg ccacgtgatg
     2701 acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag ggaagcagac
     2761 atcgatgggg atggtcaggt aaactacgaa gagtttgtac aaatgatgac agcgaagtga
     2821 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2881 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2941 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3001 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3061 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3121 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3181 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     3241 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3301 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3361 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3421 catcgatacc gtcgacccgg gcggccgctt cgagcagaca tgataagata cattgatgag
     3481 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     3541 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3601 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac
     3661 ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg taaccacgtg
     3721 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3781 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3841 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     3901 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     3961 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4021 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4081 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4141 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     4201 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4261 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     4321 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4381 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     4441 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4501 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4561 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4621 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4681 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4741 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4801 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4861 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4921 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4981 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5041 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5101 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5161 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5221 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5281 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5341 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5401 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5461 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5521 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5581 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5641 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5701 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5761 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5821 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5881 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5941 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6001 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6061 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6121 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6181 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6241 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6301 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6361 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6421 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
