LOCUS       sequence-477847-        7075 bp    DNA     circular SYN 07-JUL-2025
DEFINITION  Cre dependent Red fluorescent voltage indicator for mammalian 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7075)
  TITLE     Schnitzer lab plasmids 2025
REFERENCE   2  (bases 1 to 7075)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pAAV-EF1_-DIO-Varnam1-WPRE
FEATURES             Location/Qualifiers
     source          1..7075
                     /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    complement(1440..1473)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1524..1557)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3032..3041
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3060..3093
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3144..3177
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3202..3790
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3255..3275)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3673..3684)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3822..4298
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4021..4040)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4338..4478
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4338..4467
                     /label=AAV2 ITR
     rep_origin      4553..5008
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4640..4659)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4850..4871
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5025..5044)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5144..5166
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5204..5222)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5290..5394
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5395..5463,5464..6255)
                     /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(5613..5632)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6426..7014
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6915..6934
                     /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 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgcctcatt acacctcgtt ctcgtagcag aacacgttga tgtcgatctg gtccaggggg
     1621 atgtactcgc cctcgctggt gatcctgctc ttcttataca gctcgtccat cccgccgccc
     1681 agattagagt acttggccac ggccacctct ctctgcacca catatgtctc gttatcagac
     1741 tcctcgatgc gctccagcct gtggtccacg gcgtgcactc ctggcatctt gatattgccc
     1801 actgtcttct tggagcggta tgtggtcaca aagttacagt gcaggtgtcc tcctccatcc
     1861 accttcaggg cgatgtcggt atatcccctc aggcctccgt ctgcagggta catcatctct
     1921 gtattgggct cccagccctt ggtcttcttc tgcatcactg ggccattgct agggaagttc
     1981 acgcccctca ccttcacgtt atacaccagc tcgccatcct ccagagaggt gtcctgtgtc
     2041 acggtcacca cgccgccatc ctcgtatctt gtcaccctct cccaggtaaa gccctcgggg
     2101 aagctctgct taaagaaatc ggggatgtcg gctggatact tgatgaaggt cctgctgccg
     2161 tacatgaagc tggtggccag gatgtcaaag gcgaaaggca gtggtcctcc ctcgatcact
     2221 ttgatcctca ttgtctgcac gccctcgtat ggcctgccct ctccctctcc ggtgcacttg
     2281 aactggtgcc cattgacaga cccttccatc acgaccttca tccgcatatt ttccttaatc
     2341 agtccggacc tccacttgaa gatagtctca tgggcaatga ggcaccccat tccaaacagt
     2401 ggcttagcga gcacatccag gattcccatg agaatggcct cattgtcgac ggacagcacg
     2461 ccactaccag acccgaaagc ccacacgaca ggataaaaca cccagaggat tgacaggaag
     2521 acgagcagac tgacatagga gctatggtac tcggggcttt tcttcagatc gtctttccaa
     2581 gtggcgttat acatcccgta tgtcaggact gcctgcatca cacatccagc gataaagtac
     2641 acccatttga acttgtgttc atcctcgaag gcgcccagaa taccaaacac gatcatgaaa
     2701 atgtctgcgc cgatgaccca agaaatcatc actccgccca tcttggtcat gacgatgaga
     2761 tcgagcagga gcagtggtgt ggtcagcacc cagctaatat atcttgcgta gaccacctgc
     2821 ctttcaccat tcattgtcag gtcctggtag ttcacggcca ttgcatagta agcggtagcg
     2881 gcgatagtgc agatggcaat gttgatatag tatgcgctct tcagctctga aggccgaaag
     2941 tgcacagcca ctccaaaagc aatagcagca gcagccataa tagcaaagac aatccactgt
     3001 gcaatcatgc cggtctcggt ttccacgtca gccatggtgg cctgcagacc ggtgctagca
     3061 taacttcgta taaagtatcc tatacgaagt tatttgcctt aacccagaaa ttatcactgt
     3121 tattctttag aatggtgcaa agaataactt cgtataatgt atgctatacg aagttatgaa
     3181 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     3241 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3301 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3361 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3421 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3481 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3541 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3601 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     3661 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3721 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3781 gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc
     3841 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     3901 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     3961 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     4021 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     4081 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     4141 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     4201 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     4261 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt
     4321 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     4381 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     4441 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     4501 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     4561 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4621 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4681 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4741 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     4801 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4861 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     4921 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4981 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     5041 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     5101 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     5161 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     5221 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     5281 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5341 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5401 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5461 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5521 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5581 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5641 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5701 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5761 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5821 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5881 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5941 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6001 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6061 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6121 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6181 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6241 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6301 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6361 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6421 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6481 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6541 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6601 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6661 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6721 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6781 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6841 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6901 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6961 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7021 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
