LOCUS       sequence-242012-        7290 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Cre-dependent soma-localized C1V1(t/t) for optogenetic stimulation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7290)
  AUTHORS   Chettih SN, Harvey CD
  TITLE     Single-neuron perturbations reveal feature-specific competition in 
            V1.
  JOURNAL   Nature. 2019 Mar 6. pii: 10.1038/s41586-019-0997-6. doi: 
            10.1038/s41586-019-0997-6.
  PUBMED    30842660
REFERENCE   2  (bases 1 to 7290)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2019 Mar 6. pii: 10.1038/s41586-019-0997-6. doi: 
            10.1038/s41586-019-0997-6."
COMMENT     Sequence Label: pAAV-EF1a-DIO-C1V1(t/t)-mRuby2-KV2.1-WPRE-SV40
FEATURES             Location/Qualifiers
     source          1..7290
                     /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..1408
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1399
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1356..1376
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1439..1472)
                     /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(1523..1556)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1763..2470)
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTHM
                     ALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAVA
                     KFAGLGGGMDELYK"
     protein_bind    3522..3555
                     /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    3606..3639
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3664..4252
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3717..3737)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4135..4146)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4297..4418
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4308..4327)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4362..4381
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4553..4693
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4553..4682
                     /label=AAV2 ITR
     rep_origin      4768..5223
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4855..4874)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5065..5086
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5240..5259)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5359..5381
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5419..5437)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5505..5609
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5610..5678,5679..6470)
                     /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(5828..5847)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6641..7229
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7130..7149
                     /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 acttccactg
      541 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
      601 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
      661 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
      721 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
      781 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
      841 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
      901 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
      961 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
     1021 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
     1081 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
     1141 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     1201 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     1261 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     1321 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     1381 tttttttctt ccatttcagg tgtcgtgagg taccggatcc tctagagtcg actccggaat
     1441 aacttcgtat aggatacttt atacgaagtt atgcagaatg gtagctggat tgtagctgct
     1501 attagcaata tgaaacctct taataacttc gtatagcata cattatacga agttatggcg
     1561 cgccttagaa tctggtggct tcagggaagt ccgtggcaca gctgatgaag ctgtcaatgc
     1621 tggacatgct ccgcatgtcg atgacgccct ccgtgcgtgc gggcagaggg gccacggggc
     1681 tgggcatgct gctcatctcc agctcctctg gaggcttgct ctgcggggcc atctccttgg
     1741 tgttgaggat gggctgggac tgcttgtaca gctcgtccat cccaccacca agcccggcga
     1801 acttggcaac tgcgtgttcg cgttgtacta cgaacatttc attgtcactt tcctctaacc
     1861 tttccaggcg gtgatcaacg gcatggatac cgggcatctt gatgttcccg acggtctttt
     1921 ttgacctgta agttgttacg aaagagcaag acagatggcc accaccatca actttcagtg
     1981 ccatatgagt gtatcccctc agaccaccat ctgctggata catcatctct gtattaggct
     2041 cccaaccctt ggtcttcttc tgcatcacgg gaccattgga gggaaagttt acccctctga
     2101 cttggacgtg gtaaacgaga cagccatcct caaggctggt gtcctgcatg acggtgacga
     2161 ctccaccatc ttcgtatctc gtaactcttt cccaagtaaa accctcagga aaggactgtt
     2221 taaagaaatc aggaatgcct ttcgggtact tgataaaagt acggctgcca tacatgaacg
     2281 acgtggcaag aatgtcaaag gcaaatggca ggggtcctcc ctcgatgact ttgatcctca
     2341 tggtttgagt tcccatgtac ggattgcctt ctccttcacc tgtgcatttg aattggtggc
     2401 cgttgaccga accttccatg accaccttca tacgcatatt ttccttgatc agctcttcgc
     2461 ccttagacac ggcggccgcg tcctcctctt cagccaccag tgtctcaacc tccatctctt
     2521 ggccggcgat cgtaattttc tgcttctttc tgatgtcgcc atacaacagg atgtgctcgt
     2581 ggattttgac gcgcaggtaa ttccccagca caccccacat gttctttgca atcaggtcga
     2641 ggatggagtg cccaatggca gatccataag ggctgatatg accgaatcct tccgtgccca
     2701 gcagaaacag gactggaaac attccccaag ccacaaagaa tgtccaggcc atcactcgga
     2761 ccagttctcg acaaatccct tttggaactg tgtggaatgc ttcgatataa actttagcag
     2821 catgaaaata ggtatacata ccatagctca atgagatgag gaagaacagg atcttcgtcc
     2881 accccgtaca catggcgctg gttgcgcccc acacgatgca tcccacatca gaaaccaaca
     2941 ggcccatagt ccttttgcta taatcgtcct tcaggcctgt gaggttgctc agatggatca
     3001 gaaggaccgg gcaggtgagc agccaggtcg cgtaacgaag ccacacggtc ttgttgccgt
     3061 tggatgagta gatcaccgca ggttcgtcaa actcatggaa atactcgatg atgaacttga
     3121 tcatctcgat cgtggccacg taaatggtct cccagccgca agtagacttc caggtctggt
     3181 agccgtagaa catcaggcag agcgctgaaa gcgcaaaagt aatccactgc agaatgttgg
     3241 cagccaactt ctcggcattt gttccgttgg atttaagcca agccaagcag aagcactggc
     3301 cgttgttcgg gatgcaaata acagaaccat tgttctcaag agtgtatgag gtttggaaga
     3361 gcatgcgctc gtgggcacgg tggaaaacgt agtcggggcc atcctgagtc gcgaccggca
     3421 ccgtcgcgtc actgccagtc gaggctcctg cgctgccggc cgccagcgcc actgccagcg
     3481 ctagggcaag aagccatggc ctccgcgaca tggtggctag cataacttcg tataaagtat
     3541 cctatacgaa gttatttgcc ttaacccaga aattatcact gttattcttt agaatggtgc
     3601 aaagaataac ttcgtataat gtatgctata cgaagttatg aattcgatat caagcttatc
     3661 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3721 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3781 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3841 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3901 actggttggg gcattgccac cacctgtcag ctcctttccg gaactttcgc tttccccctc
     3961 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     4021 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     4081 ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     4141 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     4201 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     4261 cgagcgctgc tcgagagatc tttcgagcag acatgataag atacattgat gagtttggac
     4321 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     4381 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     4441 ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca
     4501 aatgtggtaa aatctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc
     4561 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     4621 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4681 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4741 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     4801 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     4861 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4921 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4981 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5041 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     5101 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     5161 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     5221 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     5281 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     5341 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     5401 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     5461 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5521 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5581 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5641 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5701 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5761 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5821 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5881 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5941 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6001 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6061 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6121 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6181 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6241 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6301 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6361 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6421 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6481 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6541 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6601 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6661 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6721 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6781 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6841 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6901 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6961 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7021 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7081 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7141 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7201 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7261 tggccttttg ctggcctttt gctcacatgt
//
