LOCUS       sequence-432142-        6569 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides AND intersectional (Cre+Flp) expression of ArchT-eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6569)
  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 6569)
  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-ArchT(eGFP)-WPRE
FEATURES             Location/Qualifiers
     source          1..6569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..42
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     236..258
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     250..267
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     251..267
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   293..422
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        456..903
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    1012..1059
                     /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)."
     protein_bind    complement(1072..1105)
                     /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(1198..1231)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(1252..1965,1966..1968))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(1255..1965,1966..1968))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1297..1318)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1646..1665
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1905..1926
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2724..2733
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2848..2895)
                     /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)."
     misc_feature    2925..3513
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2978..2998)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3396..3407)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3537..4013
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3736..3755)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4040..4180
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4040..4169
                     /label=AAV2 ITR
     primer_bind     complement(4220..4236)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4220..4236)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4233..4255)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4244..4260
                     /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)."
     promoter        complement(join(4268..4274,4275..4292,4293..4298))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4313..4334
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4622..5210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4702..4721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5381..6172,6173..6241))
                     /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     6004..6023
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6242..6346)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6414..6432
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6470..6492)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
       61 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
      121 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
      181 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
      241 tcacgacgtt gtaaaacgac ggccagtgaa ttgacgcgta ttgggatcca atcctgcagg
      301 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc
      361 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
      421 ctgcggccgc agatgctgca cgggcccgcg atcgcagtgc aagtgggttt taggaccagg
      481 atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga caagcaccca
      541 acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa caggatgcgg
      601 cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc cccgcctggc
      661 ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg
      721 caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc agccggaccg
      781 caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg cggcgccggc
      841 gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg
      901 cagggtaccg ctagcgaagt tcctattctt caaaaggtat aggaacttca tcaaaatagg
      961 aagaccaatg cttcaccatc gacccgaatt gccaagcatc accatcgaca cgaagttcct
     1021 attccgaagt tcctattctc tagaaagtat aggaacttcg tcgaccttaa gataacttcg
     1081 tatagcatac attatacgaa gttataaaaa caaaaagaaa agcgcgtttc gtcctatttg
     1141 ggactcgtca gctggatgta cgcgtttccg cgaatccagg acgcgcaaag aaagaaaata
     1201 acttcgtata gcatacatta tacgaagtta tgaattcatg catgtgcatg cttacttgta
     1261 cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg
     1321 atcgcgcttc tcgttggggt ctttgctcag ggcggactgg gtgctcaggt agtggttgtc
     1381 gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg
     1441 cacgctgccg tcctcgatgt tgtggcggat cttgaagttc accttgatgc cgttcttctg
     1501 cttgtcggcc atgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag
     1561 gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc
     1621 gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt
     1681 gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg
     1741 gtcggggtag cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca
     1801 gggcacgggc agcttgccgg tggtgcaaat gaacttcagg gtcagcttgc cgtaggtggc
     1861 atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc
     1921 gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcactg ctactaccgg
     1981 tcggggctcg ggggcctcgg tgtcgcccag gatggccctg ctcctcagca ggatgaagcc
     2041 gaagcccact ttggcggtca cgtccagcac catgaacagc agggtctcga ttcccagtcc
     2101 caccactcca gctccctcgg ttccgatgat ccacaggatg gggtaggcgg tccacagcac
     2161 cagcaccagg gcggtcaggg tgttgaaggt gctagccacc tctggtcccc tctccttagc
     2221 agcagccctc aggctggtag ccaggaagta cagcaccacg atcatgcaga tggtgctgaa
     2281 cagccaccag ctgtacctag ccagtggggt gtggctcagg gcgccgatca ggccggtcac
     2341 gatcatcagg gcgtccactc ccaccagggt tccgatgctc accctgtcca ccttagccag
     2401 cagggccagg tcgagcagaa gcaaaggggt ggtgaacagc cagtcggcgt atctggcgta
     2461 gtagatgtcc agcacctcgc cggccacggt cacctcggtc aggccgatgc cgaagaacat
     2521 gctcaggtag gcggcgctgg cgatgccggg caccaggatt gtgatgctgt agtactccct
     2581 ggcctccttg tcggtcacgc cccagccctt cacgatgaag tagaaggtgc cgatcagcat
     2641 cagcagggtt ccgattccca gccacagggt ctctggcctt ccgtcgccca gcaggtcgta
     2701 tcctgcctgc agagcgatgg ggtccatggt ggcactagta ccggtctcga ggaagttcct
     2761 ataccttttg aagaatagga acttcatcaa aataggaaga ccaatgcttc accatcgacc
     2821 cgaattgcca agcatcacca tcgacacgaa gttcctatac tttctagaga ataggaactt
     2881 cggaatagta acttccgtac gaagcttata tgcggccgat cgataatcaa cctctggatt
     2941 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3001 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3061 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3121 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3181 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3241 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3301 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     3361 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3421 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3481 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtagatcg atctacgggt
     3541 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     3601 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     3661 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     3721 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     3781 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     3841 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     3901 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     3961 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     4021 tgtaggtaac cggagctcca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     4081 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     4141 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg attggatccc aatggcgcgc
     4201 cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4261 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4321 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4381 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4441 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4501 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4561 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4621 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4681 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4741 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4801 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4861 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4921 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4981 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5041 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5101 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5161 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5221 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5281 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5341 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5401 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5461 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5521 gagaaccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5581 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5641 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5701 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5761 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5821 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5881 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5941 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6001 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6061 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6121 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6181 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6241 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6301 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6361 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     6421 gtatcacgag gcccttttgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     6481 tgcagctccc ggagactgtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     6541 gtcagggcgc gtcagcgggt gttggcggg
//
