LOCUS       sequence-349049-        6333 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV-mediated expression of GCaMP8f under the EF1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6333)
  TITLE     Looger Lab GCaMP8 plasmids
REFERENCE   2  (bases 1 to 6333)
  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"
COMMENT     Sequence Label: AAV-EF1a-jGCaMP8f-WPRE
FEATURES             Location/Qualifiers
     source          1..6333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(39..627)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(119..138)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(798..1589,1590..1658))
                     /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     1421..1440
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1659..1763)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1789..2244)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1926..1947)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2138..2157
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2316..2445
                     /label=AAV2 ITR
     promoter        2592..3769
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2823..3761
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3718..3738
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3808..3825
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     4107..4128
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4234..4255)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4495..4514)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    5086..5674
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5139..5159)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5557..5568)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5677..5693)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5678..5694)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5723..5844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5734..5753)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5788..5807
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6006..6135
                     /label=AAV2 ITR
ORIGIN
        1 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
       61 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      121 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      181 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      241 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      301 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      361 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      421 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
      481 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
      541 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
      601 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
      661 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
      721 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
      781 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
      841 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
      901 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
      961 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1021 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     1081 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     1141 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     1201 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     1261 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     1321 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     1381 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1441 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     1501 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     1561 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     1621 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     1681 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     1741 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     1801 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     1861 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     1921 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     1981 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2041 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2101 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2161 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2221 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     2281 ggcgatcggt gcgggcctct tcgctattac gccagctgcg cgctcgctcg ctcactgagg
     2341 ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
     2401 gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag ttaatgatta
     2461 acccgccatg ctacttatct acgtagccat gctctaggaa gatccaagct ttgcaaagat
     2521 ggataaagtt ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag
     2581 gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
     2641 agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa
     2701 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
     2761 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
     2821 aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg
     2881 tgccttgaat tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg
     2941 gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag
     3001 ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct
     3061 gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc
     3121 tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg
     3181 tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc
     3241 ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg
     3301 ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc
     3361 ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct
     3421 caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa
     3481 gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca
     3541 ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt
     3601 tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc
     3661 acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca
     3721 agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtgg aattcaagct
     3781 gctagcaagg atccacccgc caccatgcat catcaccatc atcacacgcg tcgcaagaag
     3841 accttcaagg aggtggccac cgccgtgaag atcatcgccc gcctgatggg cctgaagatc
     3901 aacgtctata tcaaggccga caagcagaag aacggcatca aggcgaactt ccacatccgc
     3961 cacaacatcg aggacggcgg cgtgcagctc gcctaccact accagcagaa cacccccatc
     4021 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcgtggagtc caaactttcg
     4081 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     4141 atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat ggtgagcaag
     4201 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     4261 ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc
     4321 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     4381 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     4441 ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt caaggacgac
     4501 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     4561 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     4621 aacctgccgg accaactgac tgaagagcag atcgcagaat ttaaagaggc tttctcccta
     4681 tttgacaagg acggggatgg gacaataaca accaaggagc tggggacggt gatgcggtct
     4741 ctcggactga accccacaga agcagagctg caggacatga tcaatgaagt agatgccgac
     4801 ggtgacggca caatcgactt ccctgagttc ctgacaatga tggcaagaaa aatgaaatac
     4861 agggacacgg aagaagaaat tagagaagcg ttcggtgtgt ttgataagga tggcaatggc
     4921 tacatcagtg cagcagagct tcgccacgtg atgacaaacc ttggagagaa gttaacagat
     4981 gaagaggttg atgaaatgat cagggaagca gacatcgatg gggatggtca ggtaaactac
     5041 gaagagtttg tacaaatgat gacagcgaag taaaagctta tcgataatca acctctggat
     5101 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     5161 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     5221 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     5281 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     5341 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     5401 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     5461 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     5521 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     5581 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     5641 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgacccggg
     5701 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     5761 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     5821 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     5881 cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     5941 gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa
     6001 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     6061 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     6121 cgagcgagcg cgcagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6181 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6241 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6301 gcaggaaaga acatgtgagc aaaaggccag caa
//
