LOCUS       sequence-257785-        6450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  INTRSECT 2.0.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6450)
  AUTHORS   Hafner G, Witte M, Guy J, Subhashini N, Fenno LE, Ramakrishnan C, 
            Kim YS, Deisseroth K, Callaway EM, Oberhuber M, Conzelmann KK, 
            Staiger JF
  TITLE     Mapping Brain-Wide Afferent Inputs of Parvalbumin-Expressing 
            GABAergic Neurons in Barrel Cortex Reveals Local and Long-Range 
            Circuit Motifs.
  JOURNAL   Cell Rep. 2019 Sep 24;28(13):3450-3461.e8. doi: 
            10.1016/j.celrep.2019.08.064.
  PUBMED    31553913
REFERENCE   2  (bases 1 to 6450)
  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"; doi: 
            "10.1016/j.celrep.2019.08"; journalName: "Cell Rep"; date: 
            "2019-09-24- 24"; volume: "28"; issue: "13"; pages: "3450-3461.e8"
COMMENT     Sequence Label: pAAV-nEF-Con/Fon TVA-mCherry
FEATURES             Location/Qualifiers
     source          1..6450
                     /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=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     primer_bind     complement(1101..1120)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1266..1299)
                     /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(1350..1383)
                     /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."
     primer_bind     complement(1917..1935)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    2134..2167
                     /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    2218..2251
                     /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."
     regulatory      2419..2428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2435..2468)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     misc_feature    2577..3165
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2630..2650)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3048..3059)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3197..3673
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3396..3415)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3713..3853
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3713..3842
                     /label=AAV2 ITR
     rep_origin      3928..4383
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4015..4034)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4225..4246
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4400..4419)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4519..4541
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4579..4597)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4665..4769
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4770..4838,4839..5630)
                     /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(4988..5007)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5801..6389
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6290..6309
                     /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 tcaaatagta
      721 taggaacttc gcagagtagt agctggattg tagctgctat tagcaatcta aaacctctta
      781 gaagttccta ttcttcaaaa ggtataggaa cttcggcgcg ccttacttgt acagctcgtc
      841 catgccgccg gtggagtggc ggccctcggc gcgttcgtac tgttccacga tggtgtagtc
      901 ctcgttgtgg gaggtgatgt ccaacttgat gttgacgttg taggcgccgg gcagctgcac
      961 gggcttcttg gccttgtagg tggtcttgac ctcagcgtcg tagtggccgc cgtccttcag
     1021 cttcagcctc tgcttgatct cgcccttcag ggcgccgtcc tcggggtaca tccgctcgga
     1081 ggaggcctcc cagcccatgg tcttcttctg cattacgggg ccgtcggagg ggaagttggt
     1141 gccgcgcagc ttcaccttgt agatgaactc gccgtcctgc agggaggagt cctgtgggac
     1201 gacatgactt aaccagttaa ccatgttctc tgaacatgag aagggctggc cactctccag
     1261 tcgacataac ttcgtatagg atactttata cgaagttatg cagaatggta gctggattgt
     1321 agctgctatt agcaatatga aacctcttaa taacttcgta tagtatacct tatacgaagt
     1381 tatactagtt gtgccccccc ttttttttat aaaattgtat taatgttata tacatatctc
     1441 ctgtatgtga cccatgtgct tatgactcta tttctcatgt gtttaggact ggctgtgcga
     1501 cggacacccc gactgcgacg acgggcggga cgagtggggc tgcgggacca gcgcgatccc
     1561 cgcggtgccc acggacaacg gcacagaggc tcccactgcc cctgctcctg gacgtgctct
     1621 gccagccagg aatcacggcc gcatgtggat gctgatcact gcagtgctcc tgtgctgcct
     1681 ggtagctgtg ggtggtatcg ctgcatgggg gaagtccaaa gcaaaaagca ggtctgacat
     1741 cttcagtctt gaaagcgcat ccaagggggg tggggtgagc aagggcgagg aggataacat
     1801 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     1861 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     1921 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     1981 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     2041 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     2101 gaccgtgacc caggtgagta caggaggggt accataactt cgtataaagt atcctatacg
     2161 aagttatttg ccttaaccca gaaattatca ctgttattct ttagaatggt gcaaagaata
     2221 acttcgtata aggtatacta tacgaagtta tgctagcaac cccgacacgt acctgcgggt
     2281 aacactcccc gtgggcaccg ggcggctccg agcagcggaa ctgaccgggg gggcaacggg
     2341 acaaagaacc gttaccggac ccgttaccgg tcacgttacc gggcagcagc agcagcagca
     2401 gcgcgggcag cagccgcgcc atggtggcga gctcgaagtt cctatactat ttgaagaata
     2461 ggaacttctt gccttaaccc agaaattatc actgttattc tttagaatgg tgcaaagaga
     2521 agttcctata ccttttgaag aataggaact tcgaattcga tatcaagctt atcgataatc
     2581 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2641 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2701 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2761 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2821 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     2881 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2941 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     3001 atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     3061 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     3121 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc
     3181 tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     3241 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     3301 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     3361 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     3421 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     3481 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     3541 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     3601 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     3661 cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc
     3721 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     3781 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     3841 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3901 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     3961 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     4021 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4081 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4141 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4201 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     4261 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     4321 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     4381 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     4441 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4501 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4561 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4621 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4681 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4741 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4801 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4861 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4921 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4981 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5041 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5101 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5161 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5221 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5281 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5341 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5401 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5461 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5521 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5581 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5641 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5701 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5761 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5821 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5881 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5941 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6001 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6061 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6121 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6181 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6241 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6301 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6361 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6421 tggccttttg ctggcctttt gctcacatgt
//
