LOCUS       sequence-432006-        7224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides AND intersectional (Cre+Flp) expression of N2C 
            glycoprotein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7224)
  AUTHORS   Hughes AC, Pollard BG, Xu B, Gammons JW, Chapman P, Bikoff JB, 
            Schwarz LA
  TITLE     A Novel Single Vector Intersectional AAV Strategy for Interrogating 
            Cellular Diversity and Brain Function.
  JOURNAL   bioRxiv. 2023 Feb 8:2023.02.07.527312. doi: 
            10.1101/2023.02.07.527312. Preprint.
  PUBMED    36798174
REFERENCE   2  (bases 1 to 7224)
  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: "bioRxiv. 
            2023 Feb 8:2023.02.07.527312. doi: 10.1101/2023.02.07.527312. 
            Preprint."
COMMENT     Sequence Label: pAAV-hSyn-ConVERGD-N2cG-WPRE
FEATURES             Location/Qualifiers
     source          1..7224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(106..125)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      919..1507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1408..1427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1795..1816
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1831..1836,1837..1854,1855..1861)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1869..1885
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1874..1896
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1893..1909
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1893..1909
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   1949..2089
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1960..2089
                     /label=AAV2 ITR
     polyA_signal    2116..2592
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     2374..2393
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    2616..3204
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2722..2733
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3131..3151
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    3234..3281
                     /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)."
     CDS             3406..3783
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSSK"
     CDS             3802..3891
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             3901..3957
                     /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    5553..5586
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5679..5712
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(5725..5772)
                     /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)."
     promoter        complement(5881..6328)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   6362..6491
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     complement(6517..6534)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6517..6533)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6526..6548)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6742..6761)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6861..6883
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6921..6939)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7007..7111
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7112..7180,7181..7224,1..748)
                     /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"
ORIGIN
        1 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
       61 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      121 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
      181 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
      241 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
      301 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
      361 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
      421 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
      481 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
      541 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
      601 tggttctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
      661 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
      721 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
      781 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
      841 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
      901 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
      961 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1021 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1081 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1141 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1201 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1261 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1321 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1381 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1441 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1501 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1561 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     1621 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     1681 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     1741 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     1801 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     1861 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     1921 caagctcggc gcgccattgg gatccaatcc tgcaggcagc tgcgcgctcg ctcgctcact
     1981 gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
     2041 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg gagctccggt
     2101 tacctacaaa atcagaagga cagggaaggg agcagtggtt cacgcctgta atcccagcaa
     2161 tttgggaggc caaggtgggt agatcacctg agattaggag ttggagacca gcctggccaa
     2221 tatggtgaaa ccccgtctct accaaaaaaa caaaaattag ctgagcctgg tcatgcatgc
     2281 ctggaatccc aacaactcgg gaggctgagg caggagaatc gcttgaaccc aggaggcgga
     2341 gattgcagtg agccaagatt gtgccactgc actccagctt ggttcccaat agaccccgca
     2401 ggccctacag gttgtcttcc caacttgccc cttgctccat accacccccc tccaccccat
     2461 aatattatag aaggacacct agtcagacaa aatgatgcaa cttaatttta ttaggacaag
     2521 gctggtgggc actggagtgg caacttccag ggccaggaga ggcactgggg aggggtcaca
     2581 gggatgccac ccgtagatcg atctacggta tcgatgcggg gaggcggccc aaagggagat
     2641 ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc cgcagagccg gcagcaggcc
     2701 gcgggaagga aggtccgctg gattgagggc cgaagggacg tagcagaagg acgtcccgcg
     2761 cagaatccag gtggcaacac aggcgagcag ccaaggaaag gacgatgatt tccccgacaa
     2821 caccacggaa ttgtcagtgc ccaacagccg agcccctgtc cagcagcggg caaggcaggc
     2881 ggcgatgagt tccgccgtgg caatagggag ggggaaagcg aaagtcccgg aaaggagctg
     2941 acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca gcaaacacag tgcacaccac
     3001 gccacgttgc ctgacaacgg gccacaactc ctcataaaga gacagcaacc aggatttata
     3061 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
     3121 agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca
     3181 aattttgtaa tccagaggtt gattatcgat cggccgcata taagcttcgt acggaagtta
     3241 ctattccgaa gttcctattc tctagaaagt ataggaactt cgtgtcgatg gtgatgcttg
     3301 gcaattcggg tcgatggtga agcattggtc ttcctatttt gatgaagttc ctattcttca
     3361 aaaggtatag gaacttcctc gagaccggta ctagtggccg ccaccatgcc agagccagcg
     3421 aagtctgctc ccgccccgaa aaagggctcc aagaaggcgg tgactaaggc gcagaagaaa
     3481 ggcggcaaga agcgcaagcg cagccgcaag gagagctatt ccatctatgt gtacaaggtt
     3541 ctgaagcagg tccaccctga caccggcatt tcgtccaagg ccatgggcat catgaactcg
     3601 tttgtgaacg acattttcga gcgcatcgca ggtgaggctt cccgcctggc gcattacaac
     3661 aagcgctcga ccatcacctc cagggagatc cagacggccg tgcgcctgct gctgcctggg
     3721 gagttggcca agcacgccgt gtccgagggt actaaggcca tcaccaagta caccagctct
     3781 aaggatggat ccggcgccgt ttacccatac gatgttcctg actatgcggg ctatccctat
     3841 gacgtcccgg actatgcagg atcctatcca tatgacgttc cagattacgc tggaagcgga
     3901 gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctatg
     3961 gttcctcagg ttcttttgtt tgtactcctt ctgggttttt cgttgtgttt cgggaagttc
     4021 cccatttaca cgataccaga cgaacttggt ccctggagcc ctattgacat acaccatctc
     4081 agctgtccaa ataacctggt tgtggaggat gaaggatgta ccaacctgtc cgagttctcc
     4141 tacatggaac tcaaagtggg atacatctca gccatcaaag tgaacgggtt cacttgcaca
     4201 ggtgttgtga cagaggcaga gacctacacc aactttgttg gttatgtcac aaccacattc
     4261 aagagaaagc atttccgccc caccccagac gcatgtagag ccgcgtataa ctggaagatg
     4321 gccggtgacc ccagatatga agagtcccta cacaatccat accccgacta ccactggctt
     4381 cgaactgtaa gaaccaccaa agagtccctc attatcatat ccccaagtgt gacagatttg
     4441 gacccatatg acaaatccct tcactcaagg gtcttccctg gcggaaagtg ctcaggaata
     4501 acggtgtcct ctacctactg ctcaactaac catgattaca ccatttggat gcccgagaat
     4561 ccgagaccaa ggacaccttg tgacattttt accaatagca gagggaagag agcatccaac
     4621 gggaacaaga cttgcggctt tgtggatgaa agaggcctgt ataagtctct aaaaggagca
     4681 tgcaggctca agttatgtgg agttcttgga cttagactta tggatggaac atgggtcgcg
     4741 atgcaaacat cagatgagac caaatggtgc cctccagatc agttggtgaa tttgcacgac
     4801 tttcgctcag acgagattga gcatctcgtt gtggaggagt tagtcaagaa aagagaggaa
     4861 tgtctggacg cattagagtc catcatgacc accaagtcag taagtttcag acgtctcagt
     4921 cacctgagaa aacttgtccc agggtttgga aaagcatata ccatattcaa caaaaccttg
     4981 atggaggctg atgctcacta caagtcagtc cggacctgga atgagatcat cccctcaaaa
     5041 gggtgtttga aagttggagg aaggtgccat cctcatgtga acggggtgtt tttcaatggt
     5101 ataatattag ggcctgacga ccatgtccta atcccagaga tgcaatcatc cctcctccag
     5161 caacatatgg agttgttgga atcttcagtt atccccctga tgcaccccct ggcagaccct
     5221 tctacagttt tcaaagaagg tgatgaggct gaggattttg ttgaagttca cctccccgat
     5281 gtgtacaaac agatctcagg ggttgacctg ggtctcccga actggggaaa gtatgtattg
     5341 atgactgcag gggccatgat tggcctggtg ttgatatttt ccctaatgac atggtgcaga
     5401 agagccaatc gaccagaatc gaaacaacgc agttttggag ggacaggggg gaatgtgtca
     5461 gtcacttccc aaagcggaaa agtcatacct tcatgggaat catataagag tggaggtgag
     5521 atcagactgt gagcatgcac atgcatgaat tcataacttc gtataatgta tgctatacga
     5581 agttattttc tttctttgcg cgtcctggat tcgcggaaac gcgtacatcc agctgacgag
     5641 tcccaaatag gacgaaacgc gcttttcttt ttgtttttat aacttcgtat aatgtatgct
     5701 atacgaagtt atcttaaggt cgacgaagtt cctatacttt ctagagaata ggaacttcgg
     5761 aataggaact tcgtgtcgat ggtgatgctt ggcaattcgg gtcgatggtg aagcattggt
     5821 cttcctattt tgatgaagtt cctatacctt ttgaagaata ggaacttcgc tagcggtacc
     5881 ctgcgctctc aggcacgaca cgactcctcc gctgcccacc gcagactgag gcagcgctga
     5941 gtcgccggcg ccgcagcgca gatggtcgcg cccgtgcccc cctatctcgc gcctcgcgtg
     6001 gtgcggtccg gctgggccgg cggcggcgcg gacgcgacca aggtggccgg gaaggggagt
     6061 ttgcggggga ccggcgagtg acgtcagcgc gccttcagtg ctgaggcggc ggtggcgcgc
     6121 gccgccaggc gggggcgaag gcactgtccg cggtgctgaa gctggcagtg cgcacgcgcc
     6181 tcgccgcatc ctgtttcccc tccccctctc tgatagggga tgcgcaattt ggggaatggg
     6241 ggttgggtgc ttgtccagtg ggtcggggtc ggtcgtcagg taggcacccc caccccgcct
     6301 catcctggtc ctaaaaccca cttgcactgc gatcgcgggc ccgtgcagca tctgcggccg
     6361 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     6421 gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag
     6481 ctgcctgcag gattggatcc caatacgcgt caattcactg gccgtcgttt tacaacgtcg
     6541 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     6601 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     6661 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     6721 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     6781 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     6841 cagacaagct gtgacagtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     6901 gaaacgcgcg agacaaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     6961 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     7021 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     7081 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     7141 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     7201 agtaaaagat gctgaagatc agtt
//
