LOCUS       sequence-431695-        6776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector with Ef1a promoter and LoxFAS sites for Cre-Off 
            expression of GCaMP6f (GCaMP6f will not be expressed in mammalian 
            cells that express Cre recombinase).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6776)
  AUTHORS   Meng C, Zhou J, Papaneri A, Peddada T, Xu K, Cui G
  TITLE     Spectrally Resolved Fiber Photometry for Multi-component Analysis of
            Brain Circuits.
  JOURNAL   Neuron. 2018 May 16;98(4):707-717.e4. doi: 
            10.1016/j.neuron.2018.04.012. Epub 2018 May 3.
  PUBMED    29731250
REFERENCE   2  (bases 1 to 6776)
  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"; doi: 
            "10.1016/j.neuron.2018.04.012"; journalName: "Neuron"; date: 
            "2018-05-16- 16"; volume: "98"; issue: "4"; pages: "707-717.e4"
COMMENT     Sequence Label: pAAV-Ef1a-FAS-GCaMP6f-WPRE
FEATURES             Location/Qualifiers
     source          1..6776
                     /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        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(1482..1490,1491..1508,1509..1511,1512..1544,1545..1547,
                     1548..1571,1572..1598,1599..1655,1656..1661,1662..1931,
                     1932..1949,1950..2384,2385..2390,2391..2831)
                     /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      1578..1587
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1868..1889
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1995..2016)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2256..2275)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2903..3491
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2956..2976)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3374..3385)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3523..3999
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3722..3741)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4039..4179
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4039..4168
                     /label=AAV2 ITR
     rep_origin      4254..4709
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4341..4360)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4551..4572
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4726..4745)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4845..4867
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4905..4923)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4991..5095
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5096..5164,5165..5956)
                     /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(5314..5333)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6127..6715
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6616..6635
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     1441 caacttcgta tatacctttc tatacgaagt tgtggcgcgc catgggttct catcatcatc
     1501 atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat ctgtacgacg
     1561 atgacgataa ggatctcgcc accatggtcg actcatcacg tcgtaagtgg aataagacag
     1621 gtcacgcagt cagagctata ggtcggctga gctcactcga gaacgtctat atcaaggccg
     1681 acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc gaggacggcg
     1741 gcgtgcagct cgcctaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1801 tgcccgacaa ccactacctg agcgtgcagt ccaaactttc gaaagacccc aacgagaagc
     1861 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1921 agctgtacaa gggcggtacc ggagggagca tggtgagcaa gggcgaggag ctgttcaccg
     1981 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     2041 ccggcgaggg tgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     2101 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     2161 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2221 aaggctacat ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2281 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2341 tcaaggagga cggcaacatc ctggggcaca agctggagta caacctgccg gaccaactga
     2401 ctgaagagca gatcgcagaa tttaaagagg aattctccct atttgacaag gacggggatg
     2461 ggacaataac aaccaaggag ctggggacgg tgatgcggtc tctggggcag aaccccacag
     2521 aagcagagct gcaggacatg atcaatgaag tagatgccga cggtgacggc acaatcgact
     2581 tccctgagtt cctgacaatg atggcaagaa aaatgaaata cagggacacg gaagaagaaa
     2641 ttagagaagc gttcggtgtg tttgataagg atggcaatgg ctacatcagt gcagcagagc
     2701 ttcgccacgt gatgacaaac cttggagaga agttaacaga tgaagaggtt gatgaaatga
     2761 tcagggaagc agacatcgat ggggatggtc aggtaaacta cgaagagttt gtacaaatga
     2821 tgacagcgaa ggataacggc tagcacaact tcgtatatac ctttctatac gaagttgtga
     2881 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     2941 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3001 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3061 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3121 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3181 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3241 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3301 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     3361 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3421 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3481 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     3541 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     3601 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     3661 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     3721 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     3781 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     3841 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     3901 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     3961 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     4021 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     4081 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     4141 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     4201 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     4261 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4321 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4381 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4441 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     4501 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4561 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     4621 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4681 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     4741 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4801 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4861 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4921 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4981 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5041 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5101 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5161 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5221 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5281 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5341 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5401 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5461 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5521 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5581 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5641 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5701 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5761 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5821 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5881 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5941 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6001 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6061 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6121 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6181 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6241 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6301 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6361 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6421 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6481 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6541 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6601 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6661 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6721 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
