LOCUS       sequence-207265-        6843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Can be used to express GCaMP6s in the presence of Flp recombinase. 
            Can be used to create adeno-associated virus to express 
            Flp-dependent GCaMP6s..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6843)
  TITLE     AAV GCaMP6 Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6843)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-Ef1a-fDIO-GCaMP6s
FEATURES             Location/Qualifiers
     source          1..6843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..126
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        227..1405
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          458..1396
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1353..1373
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1436..1469
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     2124..2143
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2383..2404
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2510..2531)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2744..2800)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     regulatory      2812..2821
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2828..2861)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    2970..3558
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3023..3043)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3441..3452)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3590..4066
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3789..3808)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4106..4246
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4106..4235
                     /label=AAV2 ITR
     rep_origin      4321..4776
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4408..4427)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4618..4639
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4793..4812)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4912..4934
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4972..4990)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5058..5162
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5163..5231,5232..6023)
                     /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(5381..5400)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6194..6782
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6683..6702
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
      121 gttcctgcgg ccgcacgcgt aagctttgca aagatggata aagttttaaa cagagaggaa
      181 tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct ccggtgcccg
      241 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
      301 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
      361 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
      421 cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg
      481 cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct
      541 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
      601 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
      661 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
      721 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
      781 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
      841 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
      901 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
      961 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
     1021 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
     1081 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
     1141 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
     1201 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
     1261 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
     1321 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
     1381 ttttcttcca tttcaggtgt cgtgaggtac cggatcctct agagtcgact ccggagaagt
     1441 tcctattctc tagaaagtat aggaacttcg cagaatggta gctggattgt agctgctatt
     1501 agcaatatga aacctcttag aagttcctat tcttcaaaag gtataggaac ttcggcgcgc
     1561 cttactactt cgctgtcatc atttgtacaa actcttcgta gtttacctga ccatccccat
     1621 cgatgtctgc ttccctgatc atttcatcaa cctcttcatc tgttaacttc tctccaaggt
     1681 ttgtcatcac gtggcgaagc tctgctgcac tgatgtagcc attgccatcc ttatcaaaca
     1741 caccgaacgc ttctctaatt tcttcttccg tgtccctgta tttcattttt cttgccatca
     1801 ttgtcaggaa ctcagggaag tcgattgtgc cgtcaccgtc ggcatctact tcattgatca
     1861 tgtcctgcag ctctgcttct gtggggttct gccccagaga ccgcatcacc gtccccagct
     1921 ccttggttgt tattgtccca tccccgtcct tgtcaaatag ggagaaagcc tctttaaatt
     1981 ctgcgatctg ctcttcagtc agttggtccg gcaggttgta ctccagcttg tgccccagga
     2041 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     2101 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
     2161 gctcctggat gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     2221 cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg
     2281 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
     2341 cgccctcacc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     2401 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg ctccctccgg
     2461 taccgccctt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
     2521 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttcga aagtttggac tgcacgctca
     2581 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
     2641 ggtaggcgag ctgcacgccg ccgtcctcga tgttgtggcg gatgtggaag ttcgccttga
     2701 tgccgttctt ctgcttgtcg gccttgatat agacgttctc gagtgagctc agccgaccta
     2761 tagctctgac tgcgtgacct gtcttattcc acttacgacg tgatgagtcg accatggtgg
     2821 cgctagcgaa gttcctatac tttctagaga ataggaactt cttgccttaa cccagaaatt
     2881 atcactgtta ttctttagaa tggtgcaaag agaagttcct ataccttttg aagaatagga
     2941 acttcgaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     3001 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3061 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3121 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3181 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3241 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3301 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3361 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga
     3421 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3481 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3541 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc
     3601 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     3661 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     3721 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     3781 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     3841 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     3901 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     3961 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     4021 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     4081 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     4141 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     4201 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     4261 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     4321 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4381 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4441 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4501 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4561 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4621 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     4681 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     4741 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4801 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4861 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4921 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4981 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     5041 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5101 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5161 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5221 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5281 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5341 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5401 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5461 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5521 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5581 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5641 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5701 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5761 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5821 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5881 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5941 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6001 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6061 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6121 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6181 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6241 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6301 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6361 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6421 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6481 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6541 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6601 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6661 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6721 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6781 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6841 tgt
//
