LOCUS       sequence-309461-        5861 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses FKBP12-pBirA(284-C)-HA/biochemical assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5861)
  AUTHORS   Kwak C, Shin S, Park JS, Jung M, Nhung TTM, Kang MG, Lee C, Kwon TH,
            Park SK, Mun JY, Kim JS, Rhee HW
  TITLE     Contact-ID, a tool for profiling organelle contact sites, reveals 
            regulatory proteins of mitochondrial-associated membrane formation.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 May 15. pii: 1916584117. doi: 
            10.1073/pnas.1916584117.
  PUBMED    32414919
REFERENCE   2  (bases 1 to 5861)
  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. 2020 May 15. pii: 1916584117. doi: 
            10.1073/pnas.1916584117."
COMMENT     Sequence Label: pCDNA3/FKBP12-pBirA(284-C)-HA
FEATURES             Location/Qualifiers
     source          1..5861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      39..48
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(45..47,48..368)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             492..518
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(552..570)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(553..570)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(590..607)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    596..820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      866..1294
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(953..972)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1163..1184
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1303..1323)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1308..1637
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1488..1623
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1550..1569
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1704..2498
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1758..1777)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2368..2387
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2672..2793
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2709..2728)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2763..2782
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2842..2858)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2842..2858)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2855..2877)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2866..2882
                     /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)."
     promoter        complement(join(2890..2896,2897..2914,2915..2920))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2935..2956
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3073..3090)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3244..3832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3324..3343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4003..4794,4795..4863))
                     /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     4626..4645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4864..4968)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5043..5062)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5234..5613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5614..5817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5768..5788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 taatacgact cactataggg agacccaagc ttgcggccgc caccatggga gtgcaggtgg
       61 aaaccatctc cccaggagac gggcgcacct tccccaagcg cggccagacc tgcgtggtgc
      121 actacaccgg gatgcttgaa gatggaaaga aatttgattc ctcccgggac agaaacaagc
      181 cctttaagtt tatgctaggc aagcaggagg tgatccgagg ctgggaagaa ggggttgccc
      241 agatgagtgt gggtcagaga gccaaactga ctatatctcc agattatgcc tatggtgcca
      301 ctgggcaccc aggcatcatc ccaccacatg ccactctcgt cttcgatgtg gagcttctaa
      361 aactggaagg atccgaaatc ttcggcatct ccagaggaat cgacaagcag ggagctctgc
      421 tgctggagca ggacggaatc atcaagccct ggatgggcgg agaaatctcc ctgagaagcg
      481 cagagaaggc ttacccttac gatgtaccgg attacgcata atgactcgag catgcatcta
      541 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
      601 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
      661 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
      721 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
      781 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
      841 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
      901 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
      961 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1021 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1081 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     1141 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1201 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1261 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     1321 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     1381 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     1441 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     1501 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     1561 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     1621 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     1681 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     1741 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     1801 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     1861 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     1921 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     1981 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     2041 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     2101 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     2161 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     2221 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     2281 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     2341 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     2401 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     2461 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     2521 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     2581 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     2641 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     2701 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2761 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     2821 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2881 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2941 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3001 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3061 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3121 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3181 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3241 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3361 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3421 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3481 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3541 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3601 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3661 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3721 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3781 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3901 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3961 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4021 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4081 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4141 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4201 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4261 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4321 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4381 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4441 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4501 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4561 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4621 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4681 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4741 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4801 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4861 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4921 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4981 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
     5041 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     5101 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     5161 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
     5221 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
     5281 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     5341 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     5401 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     5461 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     5521 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     5581 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     5641 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     5701 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     5761 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
     5821 tggctaacta gagaacccac tgcttactgg cttatcgaaa t
//
