LOCUS       sequence-343147-        6127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV virus for expressing TEVP and H2b-emiRFP670.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6127)
  AUTHORS   Jin L, Matsuyama M, Sullivan HA, Zhu M, Lavin TK, Hou Y, Lea NE, 
            Pruner MT, Dam Ferdinez ML, Wickersham IR
  TITLE     ""Self-inactivating"" rabies viruses are susceptible to loss of 
            their intended attenuating modification.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Feb 14;120(7):e2023481120. doi: 
            10.1073/pnas.2023481120. Epub 2023 Feb 6.
  PUBMED    37053554
REFERENCE   2  (bases 1 to 6127)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-02-14"; volume: "120(7):e2023481120. 
            doi"; pages: " 10.1073/pnas.2023481120. Epub 2023 Feb 6"
COMMENT     Sequence Label: pAAV-TREtight-H2b-emiRFP670-P2A-TEVP
FEATURES             Location/Qualifiers
     source          1..6127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    2..20
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    38..56
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    73..91
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    109..127
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    145..163
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    180..198
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    216..234
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     281..305
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             328..705
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             1651..1707
                     /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"
     misc_feature    2422..3010
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2475..2495)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2893..2904)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3018..3242
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3244..3384
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3244..3373
                     /label=AAV2 ITR
     rep_origin      3459..3914
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3546..3565)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3756..3777
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3931..3950)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4050..4072
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4110..4128)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4196..4300
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4301..4369,4370..5161)
                     /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(4519..4538)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5332..5920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5821..5840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5982..6111
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        join(6121..6127,1..308)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
ORIGIN
        1 ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacga
       61 tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg
      121 atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttat
      181 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat
      241 gtcgaggtag gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc
      301 agatcgcctg gagaattcgc cgccaccatg ccagagccag cgaagtctgc tcccgccccg
      361 aaaaagggct ccaagaaggc ggtgactaag gcgcagaaga aaggcggcaa gaagcgcaag
      421 cgcagccgca aggagagcta ttccatctat gtgtacaagg ttctgaagca ggtccaccct
      481 gacaccggca tttcgtccaa ggccatgggc atcatgaatt cgtttgtgaa cgacattttc
      541 gagcgcatcg caggtgaggc ttcccgcctg gcgcattaca acaagcgctc gaccatcacc
      601 tccagggaga tccagacggc cgtgcgcctg ctgctgcctg gggagttggc caagcacgcc
      661 gtgtccgagg gtactaaggc catcaccaag tacaccagcg ctaagggcag cggcgcggaa
      721 ggctccgtcg ccaggcagcc tgacctcttg acctgcgaac atgaagagat ccacctcgcc
      781 ggctcgatcc agccgcatgg cgcgcttctg gtcgtcagcg aacatgatca tcgcgtcatc
      841 caggccagcg ccaacgccgc ggaatttctg aatctcggaa gcgtactcgg cgttccgctc
      901 gccgagatcg acggcgatct gttgatcaag atcctgccgc atctcgatcc caccgccgaa
      961 ggcatgccgg tcgcggtgcg ctgccggatc ggcaatccct ctacggagta ctgcggtctg
     1021 atgcatcggc ctccggaagg cgggctgatc atcgaactcg aacgtgccgg cccgtcgatc
     1081 gatctgtcag gcacgctggc gccggcgctg gagcggatcc gcacggcggg ttcactgcgc
     1141 gcgctgtgcg atgacaccgt gctgctgttt cagcagtgca ccggctacga ccgggtgatg
     1201 gtgtatcgtt tcgatgagca aggccacggc ctggtattct ccgagtgcca tgtgcctggg
     1261 ctcgaatcct atttcggcaa ccgctatccg tcgtcgactg tcccgcagat ggcgcggcag
     1321 ctgtacgtgc ggcagcgcgt ccgcgtgctg gtcgacgtca cctatcagcc ggtgccgctg
     1381 gagccgcggc tgtcgccgct gaccgggcgc gatctcgaca tgtcgggctg cttcctgcgc
     1441 tcgatgtcgc cgtgccatct gcagttcctg aaggacatgg gcgtgcgcgc caccctggcg
     1501 gtgtcgctgg tggtcggcgg caagctgtgg ggcctggttg tctgtcacca ttatctgccg
     1561 cgcttcatcc gtttcgagct gcgggcgatc tgcaaacggc tcgccgaaag gatcgcgacg
     1621 cggatcaccg cgcttgagag cggaagcgga gctactaact tcagcctgct gaagcaggct
     1681 ggagacgtgg aggagaaccc tggacctatg ggcgagagcc ttttcaaggg cccgagggac
     1741 tacaacccga tctccagcac catctgtcac ctgaccaacg agagcgacgg tcacaccact
     1801 agtctgtacg gcatcggctt cggccccttc atcatcacca acaagcatct gttcaggagg
     1861 aataacggca cactgctggt gcaaagcctg cacggcgtgt tcaaagtgaa gaacacaacc
     1921 accctgcaac agcacctgat cgacggcagg gacatgatta tcatcaggat gcccaaggac
     1981 ttccccccct ttccccagaa actgaagttc agggagccac aaagggagga gcgaatctgc
     2041 ctggtgacca ccaacttcca gaccaagtcc atgagcagca tggtctctga taccagctgc
     2101 accttcccca gcagcgacgg catcttctgg aagcactgga ttcagacgaa ggatggccaa
     2161 tgcggcagcc cattggtgag cactagggac ggcttcatcg tgggcatcca cagcgccagc
     2221 aattttacca ataccaacaa ctacttcacg agcgtgccga aaaacttcat ggagctgttg
     2281 accaatcaag aggcgcagca gtgggtgagc ggctggaggc tgaacgccga cagcgttctt
     2341 tggggcggac ataaggtgtt catggtcaag cccgaggaac ccttccagcc cgttaaggaa
     2401 gccactcagc tttgagctag caatcaacct ctggattaca aaatttgtga aagattgact
     2461 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2521 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2581 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     2641 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     2701 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     2761 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     2821 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     2881 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     2941 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3001 gcctccccgc ctggatcctg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3061 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3121 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3181 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3241 ggcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3301 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     3361 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     3421 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     3481 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     3541 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3601 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3661 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3721 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3781 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     3841 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3901 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3961 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4021 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4081 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4141 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4201 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4261 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4321 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4381 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4441 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4501 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4561 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4621 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4681 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4741 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4801 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4861 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4921 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4981 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5041 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5101 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5161 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5221 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5281 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5341 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5401 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5461 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5521 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5581 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5641 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5701 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5761 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5821 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5881 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5941 tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc
     6001 tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     6061 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tacgcgtctc
     6121 gagttta
//
