LOCUS       Exported                7132 bp ds-DNA     circular SYN 03-FEB-2022
DEFINITION  Double floxed soma and AIS-localized genetically encoded voltage 
            indicator (GEVI) JEDI-2P in AAV production vector expressed under 
            the mammalian promoter (EF1a).
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-EF1a-DIO-JEDI-2P-Kv-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7132)
  TITLE     Deep, rapid, and prolonged voltage imaging with an indicator 
            optimized for two-photon microscopy
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7132)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 3, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7132
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      359..947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     848..867
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1009..1138
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        1239..2417
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1470..2408
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2365..2385
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    2448..2481
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    2532..2565
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      4105..4114
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(4125..4158)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(4209..4242)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    4267..4855
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4320..4340)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4738..4749)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4887..5363
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5086..5105)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5403..5543
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5403..5532
                     /label=AAV2 ITR
     rep_origin      5618..6073
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5705..5724)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5915..5936
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6090..6109)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6209..6231
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6269..6287)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6355..6459
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6460..188
                     /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(6678..6697)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
       61 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
      121 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
      181 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
      241 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
      301 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
      361 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
      421 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
      481 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
      541 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
      601 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
      661 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
      721 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
      781 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      841 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      901 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      961 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc
     1021 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc
     1081 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
     1141 ggccgcacgc gtaagctttg caaagatgga taaagtttta aacagagagg aatctttgca
     1201 gctaatggac cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
     1261 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg
     1321 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     1381 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     1441 ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg
     1501 gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg
     1561 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
     1621 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
     1681 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
     1741 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
     1801 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
     1861 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
     1921 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg
     1981 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
     2041 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
     2101 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
     2161 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
     2221 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
     2281 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
     2341 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
     2401 catttcaggt gtcgtgaggt accggatcct ctagagtcga ctccggaata acttcgtata
     2461 ggatacttta tacgaagtta tgcagaatgg tagctggatt gtagctgcta ttagcaatat
     2521 gaaacctctt aataacttcg tatagcatac attatacgaa gttatggcgc gccctattac
     2581 ttgtacatta tcaaaagcgg gtcgcttccg gaaaatcggt cgcgcagcta ataaagctat
     2641 caatgctgct catgctgcgc atatcaatca cgccttcggt gcgcgccggc agcggcgcca
     2701 ccgggctcgg catgctgctc atttccagtt cttccggcgg tttgctctgc ggcgccattt
     2761 ctttggtgtt cagaatcggc tggctctgac gcgtggaaga tccagatgaa cctgaagaac
     2821 ccttaagggt taccacttca agttgtttct tctgtgaagc caggcgaaat attcttatta
     2881 agataacaat tttcagaact cgaagaagag taaccatgtg cggcatctga tctgtatcat
     2941 attccagttt atgccccaga atgttgccat cttctttaaa atcggtgcct ttcagttcaa
     3001 tgcggttcac cagggtatcg ccttcaaatt tcaccaccgc gcgggtttta tatttgccat
     3061 catctttaaa gctaatggtg cgttcctgca catagccttc cggcatcgcg cttttaaaaa
     3121 aatcatggcg tttcatatga tccggatagc ggctaaagca ctgcacgcca taggttaagg
     3181 tggtcaccag ggtcggccac ggcaccggta gtttgccggt ggtgcaaata aatttcaggg
     3241 tcagtttgcc aatggtcgca tcgccttcgc cttcgccgcg cacgctaaat ttatggccgt
     3301 tcacatcgcc atccagttcc accagaatcg gcaccacgcc ggtaaacagt tcttcgccct
     3361 ggctcgcgct gccgccggtg ccgccataca gttcatccat gccgtgtgta atgcccgctg
     3421 cggtcacaaa ttccagcagc accatgtgat cgcgtttttc gttcggatct ttgctcagca
     3481 cggtctggtg gctcagataa tggttatccg gcagcagcac cgggccatcg ccaatcgggg
     3541 tgttctgctg ataatgatcc gccagctgca cgctgccatc ttccacgtta tggcgcacgg
     3601 taaagttcgc tttaatgccg tttttctgtt tatccgcggt aatatacacg ttctcgctgt
     3661 taaaggcagc aaggtcagag aaggagtagg tcatattaat tagcagagtg cccactacta
     3721 tgactgcatc caaagtaacc agtttggacc ggaaatagtt cttgaagcct tcaacaaaca
     3781 ctctcatgag aacatcaaca aggaagaaga gtgctatagc cagggaaaca ccttcaagaa
     3841 tctctctaat gcctcttttc ttctcactga tggccagatc cacaatcacc actatgatgt
     3901 ctacaatgat aagcacaact ccaaatactc tgaatccaaa tgacatcaca aacggggaaa
     3961 tttttctcct aatggtattg gaacagctgt cttgttcatt accctcatca cgaccatcat
     4021 caatctttat cgtgggctca tctgctgttg gagagctcga agttttagtg agctctgacc
     4081 cctgttcata cctcacagtc gtctccatgg tggcggtagc tagcataact tcgtataaag
     4141 tatcctatac gaagttattt gccttaaccc agaaattatc actgttattc tttagaatgg
     4201 tgcaaagaat aacttcgtat aatgtatgct atacgaagtt atgaattcga tatcaagctt
     4261 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4321 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4381 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     4441 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     4501 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     4561 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4621 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     4681 ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     4741 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     4801 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     4861 taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc
     4921 tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt
     4981 gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat
     5041 ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa
     5101 gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga
     5161 ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct
     5221 aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact
     5281 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa
     5341 ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc
     5401 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     5461 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     5521 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     5581 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     5641 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     5701 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5761 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5821 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5881 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5941 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     6001 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     6061 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     6121 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     6181 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     6241 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     6301 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     6361 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6421 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     6481 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     6541 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     6601 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6661 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6721 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6781 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6841 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6901 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6961 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     7021 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     7081 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc gg
//
