LOCUS       sequence-411126-        6104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the murine FKBP12-BirA-HA fusion protein in AAV vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6104)
  AUTHORS   Lee CS, Jung SY, Yee RSZ, Agha NH, Hong J, Chang T, Babcock LW, 
            Fleischman JD, Clayton B, Hanna AD, Ward CS, Lanza D, Hurley AE, 
            Zhang P, Wehrens XHT, Lagor WR, Rodney GG, Hamilton SL
  TITLE     Speg interactions that regulate the stability of 
            excitation-contraction coupling protein complexes in triads and 
            dyads.
  JOURNAL   Commun Biol. 2023 Sep 14;6(1):942. doi: 10.1038/s42003-023-05330-y.
  PUBMED    37709832
REFERENCE   2  (bases 1 to 6104)
  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: "Commun 
            Biol."; date: "2023-09-14"; volume: "6(1):942. doi"; pages: " 
            10.1038/s42003-023-05330-y"
COMMENT     Sequence Label: 1529_pAAV-CB-FKBP12-BirA-HA
FEATURES             Location/Qualifiers
     source          1..6104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   12..141
                     /label=AAV2 ITR
     intron          693..789
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     intron          817..949
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     932..951
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      961..970
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1303..1653,1654..1656,1657..2265)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             2269..2295
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2368..2388)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2786..2797)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2864..2881)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2870..3077
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3165..3294
                     /label=AAV2 ITR
     primer_bind     complement(3313..3330)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3313..3329)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3322..3344)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3471..3926
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3558..3577)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3768..3789
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3952..4056
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4057..4125,4126..4917)
                     /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(4275..4294)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5088..5676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5577..5596
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5964..5985
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6000..6005,6006..6023,6024..6030)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6038..6054
                     /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)."
     primer_bind     6043..6065
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6062..6078
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6062..6078
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccttaattag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt
      181 agccatgctc taggaagatc ggaattcgcc cttaagctag tatgccaagt acgcccccta
      241 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      301 actttcctac ttggcagtac atctactcga ggccacgttc tgcttcactc tccccatctc
      361 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      421 gggggcgggg ggggggggcg cgcgccaggs ggggsggggg ggggggaggg ggggggsggg
      481 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
      541 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagcg
      601 ggatcagaat gatctgatat catcgatgaa ttcgagctcg gtacccgggg atcctctaga
      661 gtcgaggaac tgaaaaacca gaaagttaac tggtaagttt agtctttttg tcttttattt
      721 caggtcccgg atccggtggt ggtgcaaatc aaagaactgc tcctcagtgg atgttgcctt
      781 tacttctagg gaagttggtc gtgaggcact gggcaggtaa gtatcaaggt tacaagacag
      841 gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt gcgtttctga
      901 taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg tgtccaggcg
      961 gccgccatgg gagtgcaggt ggagaccatc tctcctggag acgggcgcac cttcccaaag
     1021 cgcggccaga cctgcgtggt gcactacacg gggatgcttg aagatggaaa gaaatttgat
     1081 tcctctcggg acagaaacaa gccttttaag tttacactag gcaagcagga ggtgatccga
     1141 ggctgggagg aaggggtagc ccagatgagt gtgggtcaga gagccaaact gataatctcc
     1201 tcagactatg cctatggagc caccgggcac ccaggcatca tcccaccaca tgccactctt
     1261 gtttttgatg tggagcttct aaaactggaa ggaggatcgg gaatgaagga caacaccgtg
     1321 cccctgaagc tgatcgccct gctggccaac ggcgagttcc actctggcga gcagctggga
     1381 gagaccctgg gaatgagcag agccgccatc aacaagcaca tccagacact gagagactgg
     1441 ggagtggacg tgttcaccgt gcctggcaag ggctacagcc tgcctgagcc tatccagctg
     1501 ctgaacgcca agcagatcct gggacagctg gatggcggaa gcgtggccgt gctgcctgtg
     1561 atcgactcca ccaatcagta cctgctggac agaatcggag agctgaagtc cggcgacgcc
     1621 tgcatcgccg agtaccagca ggctggcaga ggaggcagag gacggaagtg gttcagccca
     1681 ttcggagcca acctgtacct gtccatgttc tggagactgg agcagggacc tgctgctgcc
     1741 atcggactga gtctggtgat cggaatcgtg atggccgagg tgctgagaaa gctgggagcc
     1801 gacaaggtga gagtgaagtg gcctaatgac ctgtacctcc aggaccgcaa gctggctggc
     1861 atcctggtgg agctgacagg caagacaggc gatgccgctc agatcgtgat cggagccgga
     1921 atcaacatgg ccatgagaag agtggaggag agcgtggtga accagggctg gatcaccctg
     1981 caggaggctg gcatcaacct ggaccggaac accctggccg ccatgctgat cagagagctg
     2041 agagccgctc tggagctgtt cgagcaggag ggactggctc cttacctgag cagatgggag
     2101 aagctggaca acttcatcaa cagacctgtg aagctgatca tcggcgacaa ggaaatcttc
     2161 ggcatctcca gaggaatcga caagcaggga gctctgctgc tggagcagga cggaatcatc
     2221 aagccctgga tgggcggaga aatctccctg agaagcgcag agaaggctta cccttacgat
     2281 gtaccggatt acgcataata ataagcttgg atccaatcaa cctctggatt acaaaatttg
     2341 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2401 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2461 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2521 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2581 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2641 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2701 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     2761 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     2821 cctgctgccg gctctgcggc ctcttccgcg tcttcgagat ctgcctcgac tgtgccttct
     2881 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2941 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3001 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3061 agcaggcatg ctggggactc gagttaaggg cgaattcccg ataaggatct tcctagagca
     3121 tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat
     3181 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3241 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagccttaa
     3301 ttaacctaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     3361 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     3421 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     3481 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3541 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3601 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3661 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3721 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3781 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3841 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3901 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     3961 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4021 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4081 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4141 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4201 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4261 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4321 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4381 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4441 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4501 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4561 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4621 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4681 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4741 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4801 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4861 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4921 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4981 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5041 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5101 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5161 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5221 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5281 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5341 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5401 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5461 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5521 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5581 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5641 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5701 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5761 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5821 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     5881 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     5941 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     6001 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     6061 acaggaaaca gctatgacca tgattacgcc agatttaatt aagg
//
