LOCUS       sequence-361482-        6476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses FKBP12-pBirA(79-C)-ha/biochemical assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6476)
  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 6476)
  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(79-C)-ha
FEATURES             Location/Qualifiers
     source          1..6476
                     /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             1107..1133
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1167..1185)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1168..1185)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1205..1222)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1211..1435
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1481..1909
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1568..1587)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1778..1799
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1918..1938)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1923..2252
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2103..2238
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2165..2184
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2319..3113
                     /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(2373..2392)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2983..3002
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3287..3408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3324..3343)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3378..3397
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3457..3473)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3457..3473)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3470..3492)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3481..3497
                     /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(3505..3511,3512..3529,3530..3535))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3550..3571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3688..3705)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3859..4447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3939..3958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4618..5409,5410..5478))
                     /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     5241..5260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5479..5583)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5658..5677)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5849..6228
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6229..6432
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6383..6403
                     /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 atccggaagc gtggccgtgc tgcctgtgat cgactccacc aatcagtacc
      421 tgctggacag aatcggagag ctgaagtccg gcgacgcctg catcgccgag taccagcagg
      481 ctggcagagg aggcagagga cggaagtggt tcagcccatt cggagccaac ctgtacctgt
      541 ccatgttctg gagactggag cagggacctg ctgctgccat cggactgagt ctggtgatcg
      601 gaatcgtgat ggccgaggtg ctgagaaagc tgggagccga caaggtgaga gtgaagtggc
      661 ctaatgacct gtacctccag gaccgcaagc tggctggcat cctggtggag ctgacaggca
      721 agacaggcga tgccgctcag atcgtgatcg gagccggaat caacatggcc atgagaagag
      781 tggaggagag cgtggtgaac cagggctgga tcaccctgca ggaggctggc atcaacctgg
      841 accggaacac cctggccgcc atgctgatca gagagctgag agccgctctg gagctgttcg
      901 agcaggaggg actggctcct tacctgagca gatgggagaa gctggacaac ttcatcaaca
      961 gacctgtgaa gctgatcatc ggcgacaagg aaatcttcgg catctccaga ggaatcgaca
     1021 agcagggagc tctgctgctg gagcaggacg gaatcatcaa gccctggatg ggcggagaaa
     1081 tctccctgag aagcgcagag aaggcttacc cttacgatgt accggattac gcataatgac
     1141 tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     1201 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1261 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1321 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1381 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     1441 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     1501 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     1561 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1621 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     1681 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     1741 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     1801 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     1861 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     1921 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     1981 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2041 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2101 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2161 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     2221 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     2281 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     2341 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     2401 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     2461 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     2521 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     2581 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     2641 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     2701 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     2761 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     2821 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     2881 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     2941 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3001 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3061 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     3121 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     3181 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3241 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     3301 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3361 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     3421 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3481 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3541 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3601 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3661 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3721 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3781 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3841 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3901 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3961 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4021 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4081 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4141 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4201 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4261 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4321 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4381 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4441 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4501 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4561 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4621 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4681 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4741 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4801 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4861 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4921 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4981 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5041 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5101 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5161 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5221 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5281 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5341 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5401 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5461 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5521 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5581 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     5641 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     5701 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     5761 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     5821 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     5881 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     5941 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     6001 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     6061 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     6121 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     6181 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     6241 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     6301 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     6361 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     6421 ataagcagag ctctctggct aactagagaa cccactgctt actggcttat cgaaat
//
