LOCUS       sequence-414425-        7424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing Voltron2 in a Flp-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7424)
  TITLE     AAV constructs for expressing genetically encoded voltage indicators
            in mice
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7424)
  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            "Unpublished"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: HT142_pAAV_EF1a_fDiO_Voltron2
FEATURES             Location/Qualifiers
     source          1..7424
                     /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"
     protein_bind    1453..1500
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      3302..3311
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3350..3397)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    3551..4139
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3604..3624)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4022..4033)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4171..4647
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4370..4389)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4687..4827
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4687..4816
                     /label=AAV2 ITR
     rep_origin      4902..5357
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4989..5008)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5199..5220
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5374..5393)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5493..5515
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5553..5571)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5639..5743
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5744..5812,5813..6604)
                     /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(5962..5981)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6775..7363
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7264..7283
                     /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 gacggctgtg
     1441 ggcccgtaac aagaagttcc tattccgaag ttcctattct ctagaaagta taggaacttc
     1501 atcaaaatag gaagaccaat gcttcaccat cgacccgaat tgccaagcat caccatcgac
     1561 agaagttcct attccgaagt tcctattctt caaaaggtat aggaacttcg cctcaaagcc
     1621 gcgtgacgag ctagttacgg cgcgccttac acctcgttct cgtagcagaa cacgttgatg
     1681 tcgatctggt ccagggggat gtactcgccc tcgctggtga tcctgctctt agtggttggc
     1741 tcgccggaaa tctcgagcgt cgacagccag cgcgcgatct cgctgccgat caggtccggg
     1801 ttgtcttctt gcagcagatt cagacccggg ccgatgtcca cagccttgca gttaggcagg
     1861 cttttggcca ggcgagcggc ttcggccggt gggatcagaa cgcctggggt gccccagaac
     1921 agcagcttcg ggacagggga ctggtgcagc cagtccatgt attcttcgac cagcgcgacg
     1981 atgttcgctg gctcaccggc gattggcagc tcgtttggga agcgccacag tggctcgcgg
     2041 tcaacaggat tcaggaacgg ctcgcggtaa tggtccatct cgacttcagt cagcgggcgg
     2101 acgacaccca tcggcagcgt accctcgata aaaacgttct gatcgatgat cagcttgcgg
     2161 ccgacgtcgg tggtgcggaa ggcctggaag gtctcgcggg caaattctgg ccattcgtcc
     2221 caggtcggga tagggcggat gaactccata aatgcaatac ctttgacgcg ctctggattg
     2281 cgcttggccc agtggaaacc cagagcggag ccccagtcgt gaatgaccag gacgacctct
     2341 tccagaccca gggcttcgat gaaggcatcc atgaagcgga cgtggtcgtc gaagaaataa
     2401 cccaggtctg gtttgtcgga tttgcccata ccgatcaggt ctggagcaat gcagcgatgg
     2461 gtcggtgcaa catgcgggat gatgttgcgc cacacgtagg aggaggtcgg gttaccgtgc
     2521 aggaacagca caggggtgcc atcgcgcgga ccaacatcga cgtagtgcat gcgctcgccc
     2581 aggacttcca cataatgggg gtcgaatgga aagccagtac cgatcttgaa gatagtctca
     2641 tgggcaatga ggcaccccat tccaaacagt ggcttagcga gcacatccag gattcccatg
     2701 agaatggcct cattgtcgac ggacagcacg ccactaccag acccgaaagc ccacacgaca
     2761 ggataaaaca cccagaggat tgacaggaag acgagcagac tgacatagga gctatggtac
     2821 tcggggcttt tcttcagatc gtctttccaa gtggcgttat acatcccgta tgtcaggact
     2881 gcctgcatca cacatccagc gataaagtac acccatttga acttgtgttc atcctcgaag
     2941 tcgcccagaa taccaaacac gatcatgaaa atgtctgcgc cgatgaccca agaaatcatc
     3001 actccgccca tcttggtcat gacgatgaga tcgagcagga gcagtggtgt ggtcagcacc
     3061 cagttaatat atcttgcgta gaccacctgc ctttcaccat tcattgtcag gtcctggtag
     3121 ttcacggcca ttgcatagta agcggtagcg gcgatagtgc agatggcaat gttgatatag
     3181 tatgcgctct tcagctctga aggccgaaag tgcacagcca ctccaaaagc aatagcagca
     3241 gcagccataa tagcaaagac aatccactgt gcaatcatgc cggtctcggt ttccacgtca
     3301 gccatggtgg cggagctagc tggcaattcg tgagcgtcgg ttgaacagcg aagttcctat
     3361 actttctaga gaataggaac ttcggaatag gaacttccgt tgggattctt cctattttga
     3421 tccaagcatc accatcgacc ctctagtcca gatctcacca tcgacccgaa gttcctatac
     3481 cttttgaaga ataggaactt cggaatagga acttcaatcc ccgcaataag ctcccatcaa
     3541 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3601 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3661 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3721 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3781 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3841 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3901 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     3961 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     4021 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     4081 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     4141 tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg
     4201 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     4261 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     4321 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     4381 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     4441 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     4501 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     4561 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     4621 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc
     4681 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     4741 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     4801 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     4861 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     4921 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     4981 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5041 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     5101 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5161 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5221 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     5281 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     5341 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     5401 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     5461 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     5521 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     5581 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     5641 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5701 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5761 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5821 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5881 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5941 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     6001 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6061 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6121 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6181 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6241 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6301 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6361 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6421 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6481 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6541 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6601 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6661 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6721 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6781 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6841 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6901 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6961 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7021 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7081 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7141 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7201 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7261 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7321 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7381 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
