LOCUS       sequence-272741-        6953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Intersectional viral expression of Arch3.3-EYFP in cells expressing 
            Cre AND NOT Flp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6953)
  AUTHORS   Fenno LE, Ramakrishnan C, Kim YS, Evans KE, Lo M, Vesuna S, Inoue M,
            Cheung KYM, Yuen E, Pichamoorthy N, Hong ASO, Deisseroth K
  TITLE     Comprehensive Dual- and Triple-Feature Intersectional Single-Vector 
            Delivery of Diverse Functional Payloads to Cells of Behaving 
            Mammals.
  JOURNAL   Neuron. 2020 Jun 20. pii: S0896-6273(20)30432-3. doi: 
            10.1016/j.neuron.2020.06.003.
  PUBMED    32574559
REFERENCE   2  (bases 1 to 6953)
  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: "Neuron. 
            2020 Jun 20. pii: S0896-6273(20)30432-3. doi: 
            10.1016/j.neuron.2020.06.003."
COMMENT     Sequence Label: pAAV-nEF-Con/Foff 2.0-Arch3.3-EYFP
FEATURES             Location/Qualifiers
     source          1..6953
                     /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        177..388
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             401..669
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     protein_bind    697..730
                     /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)"
     regulatory      823..832
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1125..1158)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    complement(1209..1242)
                     /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)."
     primer_bind     1355..1376
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1419..1475)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     protein_bind    2155..2188
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    2239..2272
                     /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)."
     primer_bind     complement(2494..2513)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2653..2904
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     2841..2862
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2938..2971)
                     /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    3080..3668
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3133..3153)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3551..3562)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3700..4176
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3899..3918)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4216..4356
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4216..4345
                     /label=AAV2 ITR
     rep_origin      4431..4886
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4518..4537)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4728..4749
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4903..4922)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5022..5044
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5082..5100)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5168..5272
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5273..5341,5342..6133)
                     /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(5491..5510)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6304..6892
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6793..6812
                     /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 gcgtaaggat ctgcgatcgc tccggtgccc gtcagtgggc
      181 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
      241 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
      301 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
      361 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
      421 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
      481 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
      541 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
      601 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
      661 ttacagatcc aagctgtgac cggcgcctac ggatccgaag ttcctattct ctagaaagta
      721 taggaacttc gcagagtagt agctggattg tagctgctat tagcaatcta aaacctctta
      781 gaagttccta ttcttcaaaa ggtataggaa cttcggcgcg ccgccaccat ggaccccatc
      841 gctctgcagg ctggttacga cctgctgggt gacggcagac ctgaaactct gtggctgggc
      901 atcggcactc tgctgatgct gattggaacc ttctactttc tggtccgcgg atggggagtc
      961 accgataagg atgcccggga atattacgct gtgactatcc tggtgcccgg aatcgcatcc
     1021 gccgcatatc tgtctatgtt ctttggtatc gggcttactg aggtgaccgt cgggggcgaa
     1081 atgttggata tctattatgc caggtaagtg tcggggttga gctcataact tcgtatagta
     1141 taccttatac gaagttattc tttgcaccat tctaaagaat aacagtgata atttctgggt
     1201 taaggcaaat aacttcgtat aggatacttt atacgaagtt atggtacccc tcctgtactc
     1261 accttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg
     1321 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
     1381 ggcaccaccc cggtgaacag ctcctcgccc ttgctcacgg gaccggggtt ttcttccacg
     1441 tctcctgctt gctttaacag agagaagttc gtggctccgg agccgggcgc gcccacgttg
     1501 atgtcgatct ggtccagggg gatgtactcg ccctcgctgg tgatcctgct cttgtcggcg
     1561 gcactgacat cggcaccggc actgggttct ggtgcctcgg tgtcgcccag aatagcccgg
     1621 gatctcaaca ggataaagcc aaagccgacc ttggcagtca cgtccaacac cataaacagc
     1681 agagtttcga tgcccaggcc caccacgcca gcgccctcag tgcctatgat ccacaggata
     1741 gggtaagcgg tccacagcac caagaccaga gctgtcaggg tgttaaaggt agatgccacc
     1801 tcggggcccc gctcctttgc agcagatcgc agggatgtag ccagaaaata gagcaccact
     1861 atcatgcaaa ttgtagagaa caaccaccaa ctgtatctgg ctatggccgt gtggctcaag
     1921 gctccgatga ggccagtgac gatcatcagg gcgtccacac ccaccagggt gccgatggtc
     1981 acccgatcca ccttagcgag aagggccaga tccagcagca gaagtggggt ggtaaacagc
     2041 cagtcggcgt acctaaacac atgagaaata gagtcataag cacatgggtc acatacagga
     2101 gattgtatat aacattaata caattttata aaaaaaaggg ggggcacaac tagtataact
     2161 tcgtataagg tatactatac gaagttatta agaggtttca tattgctaat agcagctaca
     2221 atccagctac cattctgcat aacttcgtat aaagtatcct atacgaagtt atgtcgactg
     2281 gagagtggcc agcccttctc atgttcagag aacatggtta actggttaag tcatgtcgtc
     2341 ccacagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgcagtgc
     2401 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     2461 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     2521 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     2581 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     2641 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     2701 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     2761 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac
     2821 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     2881 ctcggcatgg acgagctgta caagttctgc tacgagaacg aggtgtaatg agctagcgaa
     2941 gttcctatac tttctagaga ataggaactt ctaagcagac aatcttgtaa tacatcagca
     3001 gctagtagat tgctccagct agaagttcct ataccttttg aagaatagga acttcgaatt
     3061 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3121 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3181 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3241 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3301 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3361 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3421 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     3481 gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg
     3541 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3601 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3661 ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc
     3721 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     3781 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     3841 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     3901 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     3961 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     4021 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     4081 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     4141 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc
     4201 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     4261 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     4321 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     4381 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     4441 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4501 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4561 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4621 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     4681 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4741 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     4801 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4861 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     4921 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     4981 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5041 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     5101 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     5161 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5221 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5281 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5341 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5401 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5461 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5521 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5581 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5641 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5701 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5761 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5821 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5881 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5941 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6001 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6061 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6121 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6181 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6241 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6301 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6361 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6421 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6481 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6541 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6601 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6661 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6721 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6781 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6841 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6901 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
