LOCUS       sequence-411127-        6496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control for FKBP12-BirA construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6496)
  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 6496)
  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: 1533_pAAV-CB-AcGFP-BirA-HA
FEATURES             Location/Qualifiers
     source          1..6496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   12..141
                     /label=AAV2 ITR
     intron          692..788
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     intron          816..948
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     931..950
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      960..969
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             966..1682
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     CDS             join(1695..2045,2046..2048,2049..2657)
                     /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             2661..2687
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2760..2780)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3178..3189)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3256..3273)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3262..3469
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3557..3686
                     /label=AAV2 ITR
     primer_bind     complement(3705..3722)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3705..3721)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3714..3736)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3863..4318
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3950..3969)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4160..4181
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4344..4448
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4449..4517,4518..5309)
                     /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(4667..4686)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5480..6068
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5969..5988
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6356..6377
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6392..6397,6398..6415,6416..6422)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6430..6446
                     /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     6435..6457
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6454..6470
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6454..6470
                     /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 gggggggcgc gcgccaggcg gggcggggsg gggggggggg gggggsgggg
      481 cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta
      541 tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagcgg
      601 gatcagaatg atctgatatc atcgatgaat tcgagctcgg tacccgggga tcctctagag
      661 tcgaggaact gaaaaaccag aaagttaact ggtaagttta gtctttttgt cttttatttc
      721 aggtcccgga tccggtggtg gtgcaaatca aagaactgct cctcagtgga tgttgccttt
      781 acttctaggg aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt acaagacagg
      841 tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat
      901 aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt gtccaggcgg
      961 ccgccatggt gagcaagggc gccgagctgt tcaccggcat cgtgcccatc ctgatcgagc
     1021 tgaatggcga tgtgaatggc cacaagttca gcgtgagcgg cgagggcgag ggcgatgcca
     1081 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgcct gtgccctggc
     1141 ccaccctggt gaccaccctg agctacggcg tgcagtgctt ctcacgctac cccgatcaca
     1201 tgaagcagca cgacttcttc aagagcgcca tgcctgaggg ctacatccag gagcgcacca
     1261 tcttcttcga ggatgacggc aactacaagt cgcgcgccga ggtgaagttc gagggcgata
     1321 ccctggtgaa tcgcatcgag ctgaccggca ccgatttcaa ggaggatggc aacatcctgg
     1381 gcaataagat ggagtacaac tacaacgccc acaatgtgta catcatgacc gacaaggcca
     1441 agaatggcat caaggtgaac ttcaagatcc gccacaacat cgaggatggc agcgtgcagc
     1501 tggccgacca ctaccagcag aataccccca tcggcgatgg ccctgtgctg ctgcccgata
     1561 accactacct gtccacccag agcgccctgt ccaaggaccc caacgagaag cgcgatcaca
     1621 tgatctactt cggcttcgtg accgccgccg ccatcaccca cggcatggat gagctgtaca
     1681 agggaggatc gggaatgaag gacaacaccg tgcccctgaa gctgatcgcc ctgctggcca
     1741 acggcgagtt ccactctggc gagcagctgg gagagaccct gggaatgagc agagccgcca
     1801 tcaacaagca catccagaca ctgagagact ggggagtgga cgtgttcacc gtgcctggca
     1861 agggctacag cctgcctgag cctatccagc tgctgaacgc caagcagatc ctgggacagc
     1921 tggatggcgg aagcgtggcc gtgctgcctg tgatcgactc caccaatcag tacctgctgg
     1981 acagaatcgg agagctgaag tccggcgacg cctgcatcgc cgagtaccag caggctggca
     2041 gaggaggcag aggacggaag tggttcagcc cattcggagc caacctgtac ctgtccatgt
     2101 tctggagact ggagcaggga cctgctgctg ccatcggact gagtctggtg atcggaatcg
     2161 tgatggccga ggtgctgaga aagctgggag ccgacaaggt gagagtgaag tggcctaatg
     2221 acctgtacct ccaggaccgc aagctggctg gcatcctggt ggagctgaca ggcaagacag
     2281 gcgatgccgc tcagatcgtg atcggagccg gaatcaacat ggccatgaga agagtggagg
     2341 agagcgtggt gaaccagggc tggatcaccc tgcaggaggc tggcatcaac ctggaccgga
     2401 acaccctggc cgccatgctg atcagagagc tgagagccgc tctggagctg ttcgagcagg
     2461 agggactggc tccttacctg agcagatggg agaagctgga caacttcatc aacagacctg
     2521 tgaagctgat catcggcgac aaggaaatct tcggcatctc cagaggaatc gacaagcagg
     2581 gagctctgct gctggagcag gacggaatca tcaagccctg gatgggcgga gaaatctccc
     2641 tgagaagcgc agagaaggct tacccttacg atgtaccgga ttacgcataa taataagctt
     2701 ggatccaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     2761 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     2821 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     2881 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     2941 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     3001 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3061 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     3121 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     3181 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     3241 cgtcttcgag atctgcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3301 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3361 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3421 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggac tcgagttaag
     3481 ggcgaattcc cgataaggat cttcctagag catggctacg tagataagta gcatggcggg
     3541 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3601 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3661 gcctcagtga gcgagcgagc gcgcagcctt aattaaccta attcactggc cgtcgtttta
     3721 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     3781 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3841 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3901 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3961 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4021 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4081 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4141 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4201 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4261 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     4321 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4381 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4441 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4501 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4561 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4621 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4681 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4741 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4801 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4861 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4921 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4981 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5041 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5101 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5161 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5221 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5281 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5341 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5401 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5461 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5521 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5581 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5641 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5701 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5761 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5821 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5881 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5941 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6001 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6061 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6121 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6181 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6241 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     6301 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     6361 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     6421 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     6481 ccagatttaa ttaagg
//
