LOCUS       sequence-418041-        7581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for mapping synaptic projections. GABAergic 
            neuron-specific Dlx promoter expresses EGFP-Synaptobrevin-2 to label
            synaptic terminals and Palm-tdTomato to label neuronal soma and 
            axons..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7581)
  AUTHORS   Du W, Li E, Guo J, Arano R, Kim Y, Chen YT, Thompson A, Oh SJ, 
            Samuel A, Li Y, Oyibo HK, Xu W
  TITLE     Directed stepwise tracing of polysynaptic neuronal circuits with 
            replication-deficient pseudorabies virus.
  JOURNAL   Cell Rep Methods. 2023 Jun 16;3(6):100506. doi: 
            10.1016/j.crmeth.2023.100506. eCollection 2023 Jun 26.
  PUBMED    37426757
REFERENCE   2  (bases 1 to 7581)
  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: "Cell Rep 
            Methods."; date: "2023-06-16"; volume: "3(6):100506. doi"; pages: " 
            10.1016/j.crmeth.2023.100506. eCollection 2023 Jun 26"
COMMENT     Sequence Label: AAV-Dlx-SynaptoTAG
FEATURES             Location/Qualifiers
     source          1..7581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      23..611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     512..531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   673..802
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          1522..1654
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1637..1656
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1767..3194
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             3204..3260
                     /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"
     CDS             join(3267..3269,3270..3272,3273..3983)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3267..3269,3270..3272,3273..3983)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3312..3333)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3573..3592)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3920..3941
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4380..4968
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4433..4453)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4851..4862)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5000..5476
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5199..5218)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5516..5656
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5516..5645
                     /label=AAV2 ITR
     rep_origin      5731..6186
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5818..5837)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6028..6049
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6203..6222)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6322..6344
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6382..6400)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6468..6572
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6573..6641,6642..7433)
                     /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(6791..6810)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
       61 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
      181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      661 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt
      721 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      781 aactccatca ctaggggttc ctgcggccgc acgcgtttat ttcagaatgt tatgcactca
      841 cagtggtttg gcatgcatct ggtgaatttt ttttaacgaa aaattagtgt tggtttcgat
      901 gtatggtagc attctcccta acgtaatttg aataattcag caaagcccca ctaccagctg
      961 tacttctgca gcctcttcca ttcttttcag cattataatt ttggttaatt ttcaatttta
     1021 ggtcctacgt ctctgcaatt tgtgtatgaa taacagaata atttccctct tttgtttcgc
     1081 ctttcctgtt cctgaatcta aataaagatg gctttttagt attaaaagtg gaagaaaatt
     1141 acaggtaatt atctttgacg gtaaaaacgc tgtaatcagc gggctacatg aaaaattact
     1201 ctaattatgg ctgcatttaa gagaatggaa aaaaaccttc ttgtggataa aaaccttaaa
     1261 ttgtccccaa tgtctgcttc aaattggatg gcactgcagc tggaggcttt gttcagaatt
     1321 gatcctgggg agctacgaac ccaaagtttc acagtaggaa gcttgatatc gaattcctgc
     1381 agcccgggct gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctagc
     1441 ctgcaggtcg aggagcgcag ccttccagaa gcagagcgcg gcgccttaag ctgcagaagt
     1501 tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta aggagaccaa
     1561 tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt
     1621 cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccagtt caattacagc
     1681 tcttaagaaa gaccggtgcc accatgctgt gctgtatgag aagaaccaaa caggttgaaa
     1741 agaatgatga ggaccaaaag atcgagatgg tgagcaaggg cgaggaggtc atcaaagagt
     1801 tcatgcgctt caaggtgcgc atggagggct ccatgaacgg ccacgagttc gagatcgagg
     1861 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     1921 gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcatgtac ggctccaagg
     1981 cgtacgtgaa gcaccccgcc gacatccccg attacaagaa gctgtccttc cccgagggct
     2041 tcaagtggga gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg acccaggact
     2101 cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc
     2161 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt
     2221 acccccgcga cggcgtgctg aagggcgaga tccaccaggc cctgaagctg aaggacggcg
     2281 gccactacct ggtggagttc aagaccatct acatggccaa gaagcccgtg caactgcccg
     2341 gctactacta cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg
     2401 tggaacagta cgagcgctcc gagggccgcc accacctgtt cctggggcat ggcaccggca
     2461 gcaccggcag cggcagctcc ggcaccgcct cctccgagga caacaacatg gccgtcatca
     2521 aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga
     2581 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     2641 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct
     2701 ccaaggcgta cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg
     2761 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc
     2821 aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact
     2881 tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc
     2941 gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg
     3001 acggcggcca ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac
     3061 tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca
     3121 ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg tacggcatgg
     3181 acgagctgta caagggaagc ggagctacta acttcagcct gctgaagcag gctggagacg
     3241 tggaggagaa ccctggacct caattcatgg tgagcaaggg cgaggagctg ttcaccgggg
     3301 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     3361 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     3421 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     3481 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     3541 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     3601 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     3661 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     3721 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     3781 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     3841 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     3901 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     3961 tcggcatgga cgagctgtac aagggtggag gtggatcggc taccgctgcc accgtcccgc
     4021 ctgccgcccc ggccggcgag ggtggccccc ctgcacctcc tccaaatctt accagtaaca
     4081 ggagactgca gcagacccag gcccaggtgg atgaggtggt ggacatcatg agggtgaatg
     4141 tggacaaggt cctggagcga gaccagaagc tatcggaact ggatgatcgc gcagatgccc
     4201 tccaggcagg ggcctcccag tttgaaacaa gtgcagccaa gctcaagcgc aaatactggt
     4261 ggaaaaacct caagatgatg atcatcttgg gagtgatttg cgccatcatc ctcatcatca
     4321 tcatcgttta cttcagcact taagaattga gatctgaatt cgatatcaag cttatcgata
     4381 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     4441 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     4501 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     4561 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     4621 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     4681 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     4741 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     4801 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4861 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     4921 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     4981 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     5041 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     5101 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     5161 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     5221 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     5281 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     5341 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     5401 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     5461 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa
     5521 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     5581 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     5641 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     5701 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     5761 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     5821 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5881 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5941 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     6001 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     6061 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     6121 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     6181 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     6241 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     6301 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     6361 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     6421 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     6481 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     6541 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     6601 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     6661 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     6721 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6781 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6841 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6901 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6961 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     7021 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     7081 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     7141 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     7201 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     7261 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     7321 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     7381 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     7441 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     7501 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     7561 tccactgagc gtcagacccc g
//
