LOCUS       sequence-92026-e        6071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses splitTVA800.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6071)
  TITLE     Genetic Neuroengineering Group Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6071)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-CAG-FLEX-splitTVA800
FEATURES             Location/Qualifiers
     source          1..6071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        145..524
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        526..803
                     /label=chicken beta-actin promoter
     intron          805..1822
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1746..1768
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1830..1849
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1884..1893
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1899..1932
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1983..2016
                     /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(2393..2426)
                     /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(2477..2510)
                     /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)."
     misc_feature    2512..3100
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2565..2585)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2983..2994)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3108..3332
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3334..3474
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3334..3463
                     /label=AAV2 ITR
     rep_origin      3549..4004
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3636..3655)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3846..3867
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4021..4040)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4140..4162
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4200..4218)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4286..4390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4391..4459,4460..5251)
                     /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(4609..4628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5422..6010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5911..5930
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgtcgacatt gattattgac tagttattaa tagtaatcaa
      181 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      241 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      301 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      361 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      421 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      481 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
      541 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
      601 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccaggcg
      661 gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc
      721 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
      781 aaaagcgaag cgcgcggcgg gcggggagtc gctgcgacgc tgccttcgcc ccgtgccccg
      841 ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt
      901 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct
      961 tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg
     1021 ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc
     1081 cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag
     1141 tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg
     1201 gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc
     1261 ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc
     1321 gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg
     1381 gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg
     1441 gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct
     1501 tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc
     1561 gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc
     1621 cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct
     1681 ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag
     1741 ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca
     1801 tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca
     1861 ttttggcaaa gaattccgga gccgccacca tggttaatat aacttcgtat aatgtatgct
     1921 atacgaagtt atgcagaatg gtagctggat tgtagctgct attagcaata tgaaacctct
     1981 taataacttc gtataaagta tcctatacga agttatggcg cgccctatca gtcccatctc
     2041 accagctcac agcaaaagat ccctgcagtg atcagcatcc acatgcggcc gtgattcctg
     2101 gctggcagag cacgtccagg agcagggaca gtgggagcct ctgtgccgtt gtccgtgggc
     2161 accgcggggg tcgcgctggt cccgcagccc cactcgtccc gcccgtcgtc gcagtcgggg
     2221 tgtccgtcgc acagccagtc ctgcgggtaa cactccccgt gggcaccggg cggctccgag
     2281 cagcggaact gaccgggggg gcaacgggac aaagaaccgt taccggaccc gttaccggtc
     2341 acgttaccgg gcagcagcag cagcagcagc gcgggcagca gccgcgcacc taataacttc
     2401 gtatagcata cattatacga agttatttgc cttaacccag aaattatcac tgttattctt
     2461 tagaatggtg caaagaataa cttcgtatag gatactttat acgaagttat gaatcaacct
     2521 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2581 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2641 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2701 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2761 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2821 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2881 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     2941 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3001 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3061 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggatcctg tgccttctag
     3121 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3181 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3241 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3301 caggcatgct ggggatgcgg tgggctctat ggcaggaacc cctagtgatg gagttggcca
     3361 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3421 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     3481 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     3541 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3601 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3661 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3721 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     3781 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3841 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt
     3901 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3961 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4021 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4081 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4141 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4201 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4261 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4321 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4381 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4441 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4501 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4561 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4621 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4681 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4741 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4801 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4861 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4921 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4981 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5041 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5101 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5161 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5221 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5281 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5341 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5401 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5461 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5521 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     5581 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5641 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5701 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5761 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5821 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5881 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5941 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6001 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6061 tgctcacatg t
//
