LOCUS       sequence-496865-        5946 bp    DNA     circular SYN 12-JAN-2026
DEFINITION  CytoTape-vivo timestamp monomer for encoding time along the 
            CytoTape-vivo fiber for mouse brain in vivo applications.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5946)
  AUTHORS   Zheng L, Yan Y, Zhou B, Lim J, Shi D, An B, Ko B, Klyder E, 
            Pitchiaya S, Cai DJ, Boyden ES, Wei D, Lio P, Linghu C
  TITLE     Multiplexed, scalable analog recording of gene regulation dynamics 
            over weeks using intracellular protein tapes.
  JOURNAL   bioRxiv [Preprint]. 2025 May 10:2025.05.10.653182. doi: 
            10.1101/2025.05.10.653182.
  PUBMED    40654887
REFERENCE   2  (bases 1 to 5946)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 12, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 May 10:2025.05.10.653182. doi: 
            10.1101/2025.05.10.653182."
COMMENT     Sequence Label: pAAV-TRE3G-XRI-FLAG
FEATURES             Location/Qualifiers
     source          1..5946
                     /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
     promoter        178..556
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    186..204
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    222..240
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    258..276
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    294..312
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    330..348
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    366..384
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    402..420
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     465..489
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1833..1856
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2984..3001)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2990..3197
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3220..3349
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3411..3999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3491..3510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4170..4961,4962..5030))
                     /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     4793..4812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5031..5135)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5203..5221
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5259..5281)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5381..5400
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5417..5872)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5554..5575)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5766..5785
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgaaacaa ttctgcagga attcctcgag
      181 tttactccct atcagtgata gagaacgtat gaagagttta ctccctatca gtgatagaga
      241 acgtatgcag actttactcc ctatcagtga tagagaacgt ataaggagtt tactccctat
      301 cagtgataga gaacgtatga ccagtttact ccctatcagt gatagagaac gtatctacag
      361 tttactccct atcagtgata gagaacgtat atccagttta ctccctatca gtgatagaga
      421 acgtataagc tttaggcgtg tacggtgggc gcctataaaa gcagagctcg tttagtgaac
      481 cgtcagatcg cctggagcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac
      541 ttcctaccct cgtaaagtcg acgagctcgg tacccgggct agccaccatg attgactata
      601 ctgctgctgg tttcaccctc ctccaaggtg cacatctgta cgctcctgaa gacagaggta
      661 tctgcgatgt gctggttgcc aacggtaaga tcatcgctgt ggcctccaac attcctagcg
      721 acatcgttcc aaactgcacc gttgtcgatc tgagcggaca gatcctctgt ccaggattca
      781 ttgatcagca cgtgcacctc attggaggag gaggcgaagc tggtcctact actagaactc
      841 ccgaagtcgc actgtctcgc ctcacagagg caggcgttac ctccgtcgtg ggactcctgg
      901 gtactgatag catctctagg caccctgagt ctctgctggc aaagaccaga gccctcaatg
      961 aggagggtat ctccgcttgg atgctgactg gtgcttatca cgtcccatct cgcaccatta
     1021 ctggcagcgt ggagaaggac gttgcaatca tcgacagagt gatcggtgtt aagtgcgcta
     1081 tctccgacca caggagcgct gctcccgacg tctatcacct ggccaacatg gcagccgaga
     1141 gcagagtcgg tggtctgctc ggtggcaagc caggagttac cgtgttccac atgggtgact
     1201 ctaagaaggc actgcaaccc atctatgacc tgctggagaa ctgtgacgtc ccaatctcca
     1261 agctgctgcc tacccacgtc aacaggaacg tgcctctctt ctaccaagct ctggagttcg
     1321 caagaaaggg tggcaccatc gacatcacct ctagcattga tgaacccgtc gctccagctg
     1381 agggaatcgc aagggctgtc caagcaggta tcccactggc aagggtgaca ctcagctctg
     1441 atggcaacgg ctcccagcca ttcttcgatg atgaaggaaa tctgactcac attggcgtgg
     1501 ctggatttga aactctcctg gaaacagttc aggttctggt taaggactac gacttcagca
     1561 tttctgacgc tctcagacca ctcacctcta gcgtcgctgg cttcctgaac ctgactggta
     1621 aaggcgaaat cctccctggt aatgacgctg atctgctggt gatgacaccc gaactgcgca
     1681 ttgagcaggt ctatgcaaga ggaaagctga tggttaagga cggaaaggct tgtgtcaagg
     1741 gcacattcga gactgctggt ggctctggag gcaccggagg gtctggaggg actggaggct
     1801 ctggaggtac tggcggttct ggaggtaccg gtgactacaa ggacgatgat gacaaaggat
     1861 ccggtaagat tgaagagggc aaactggtta tctggatcaa tggcgataag ggctacaatg
     1921 gcctggcaga ggtcggtaag aagttcgaga aagatacagg catcaaagtt actgtggaac
     1981 acccagataa gctggaggag aagttccctc aggtcgcagc aactggcgac ggtccagaca
     2041 tcatcttctg ggcacacgat aggttcggtg gctatgccca atctggcctg ctggctgaga
     2101 ttactcctga taaggctttc caggacaagc tgtatccatt cacctgggat gctgtgagat
     2161 acaatggcaa gctgatcgca taccctattg ctgtggaagc tctgagcctg atctacaaca
     2221 aggacctgct gcctaatcct cctaagacat gggaagaaat ccctgcactg gacaaggaac
     2281 tgaaggccaa aggcaagtcc gcactgatgt tcaacctgca ggagccttac tttacctggc
     2341 cactgattgc tgccgacggt ggttacgctt tcaagtacga gaatggtaag tacgacatca
     2401 aagatgtggg tgtggacaat gctggtgcca aagctggtct gactttcctg gtggatctga
     2461 tcaagaacaa gcacatgaat gcagacactg actattctat cgcagaggct gctttcaaca
     2521 aaggcgaaac cgcaatgact atcaatggtc cttgggcatg gtctaacatc gacactagca
     2581 aagtcaacta cggtgtcacc gttctgccaa ccttcaaggg tcagccaagc aaacctttcg
     2641 ttggcgtgct gagcgcaggt atcaacgctg cctctcctaa caaagagctg gctaaggagt
     2701 ttctggagaa ctacctgctg acagacgaag gtctggaggc agtgaacaag gacaagccac
     2761 tgggtgccgt ggctctgaag agctacgaag aagaactggc caaggaccct cgcatcgctg
     2821 ctacaatgga gaacgcacag aagggtgaga tcatgccaaa catcccacag atgagcgcat
     2881 tctggtatgc cgtgaggacc gctgttatca acgcagcttc tggcagacag accgtggatg
     2941 aagccctgaa agacgcacag acctaactcg agagatcgat ctgcctcgac tgtgccttct
     3001 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3061 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3121 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3181 agcaggcatg ctggggacac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     3241 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     3301 gacgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcagga catgtgagca
     3361 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3421 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3481 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3541 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3601 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3661 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3721 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3781 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3841 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3901 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3961 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4021 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4081 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4141 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4201 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4261 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4321 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4381 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4441 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4501 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4561 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4621 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4681 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4741 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4801 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4861 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4921 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4981 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5041 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5101 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5161 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5221 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5281 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5341 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     5401 ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     5461 tgttaaatca gctcattttt taaccaatag accgaaatcg gcaaaatccc ttataaatca
     5521 aaagaatagc ccgagataga gttgagtgtt gttccagttt ggaacaagag tccactatta
     5581 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     5641 cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     5701 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     5761 aaggaaggga agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg
     5821 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt
     5881 tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     5941 ggcgcc
//
