LOCUS       sequence-332029-        7364 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Encodes FKBP tagged with the C-terminal fragment of split-miniSOG 
            (mSOG-Ja95-140)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7364)
  AUTHORS   Boassa D, Lemieux SP, Lev-Ram V, Hu J, Xiong Q, Phan S, Mackey M, 
            Ramachandra R, Peace RE, Adams SR, Ellisman MH, Ngo JT
  TITLE     Split-miniSOG for Spatially Detecting Intracellular Protein-Protein 
            Interactions by Correlated Light and Electron Microscopy.
  JOURNAL   Cell Chem Biol. 2019 Oct 17;26(10):1407-1416.e5. doi: 
            10.1016/j.chembiol.2019.07.007. Epub 2019 Aug 1.
  PUBMED    31378710
REFERENCE   2  (bases 1 to 7364)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.chembiol.2019.07.007"; journalName: "Cell Chem Biol"; 
            date: "2019-10-17- 17"; volume: "26"; issue: "10"; pages: 
            "1407-1416.e5"
COMMENT     Sequence Label: pcDNA3-FKBP-C-mSOG-J-alpha-IRES-mCherry
FEATURES             Location/Qualifiers
     source          1..7364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             698..1018
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1810..2520
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1955..1973)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2223..2242
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2552..2569)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2558..2782
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2828..3256
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2915..2934)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3125..3146
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3265..3285)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3270..3599
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3450..3585
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3512..3531
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3648..4673
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3648..4673
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4803..4924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4840..4859)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4894..4913
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4973..4989)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4973..4989)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4986..5008)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4997..5013
                     /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(5021..5027,5028..5045,5046..5051))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5066..5087
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5204..5221)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5375..5963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5455..5474)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6134..6925,6926..6994))
                     /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     6757..6776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6995..7099)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7174..7193)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttgccaccat gagcagggga gtacaggtag agacgatatc
      721 tcccggtgac ggacgaacat ttcccaaacg cggacagaca tgtgtcgtgc actacacagg
      781 gatgctggaa gatgggaaga aatttgatag cagccgggat cgcaacaagc cgttcaagtt
      841 catgctgggc aagcaggaag ttatcagggg atgggaggaa ggcgtggcac agatgtccgt
      901 gggacaacgc gcaaagctga caatctcacc cgactacgct tatggcgcga ccggacatcc
      961 cggaattatt cctcctcacg ccacgctggt ttttgatgtt gagctgctga aactggaatc
     1021 cgggagaggg tcctacgtta gtggtcggtc tattgccacc ggcagtggga gcggagaact
     1081 gcagtatttt attggagtgc agctcgacgg gtccgatcac gtagaaccac tgcaaaacag
     1141 gctctcagag aggacggaga tgcagagtag taagctcgtg aaagctaccg ccaccaacgt
     1201 cgatgaggcc gtctgactcg agtactccgg tattgcggta cccttgtacg cctgttttat
     1261 actcccttcc cgtaacttag acgcacaaaa ccaagttcaa tagaaggggg tacaaaccag
     1321 taccaccacg aacaagcact tctgtttccc cggtgatgtc gtatagactg cttgcgtggt
     1381 tgaaagcgac ggatccgtta tccgcttatg tacttcgaga agcccagtac cacctcggaa
     1441 tcttcgatgc gttgcgctca gcactcaacc ccagagtgta gcttaggctg atgagtctgg
     1501 acatccctca ccggtgacgg tggtccaggc tgcgttggcg gcctacctat ggctaacgcc
     1561 atgggacgct agttgtgaac aaggtgtgaa gagcctattg agctacataa gaatcctccg
     1621 gcccctgaat gcggctaatc ccaacctcgg agcaggtggt cacaaaccag tgattggcct
     1681 gtcgtaacgc gcaagtccgt ggcggaaccg actactttgg gtgtccgtgt ttccttttat
     1741 tttattgtgg ctgcttatgg tgacaatcac agattgttat cataaagcga attggatagg
     1801 atccgtacga tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     1861 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     1921 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     1981 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     2041 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     2101 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     2161 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     2221 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     2281 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     2341 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     2401 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     2461 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
     2521 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     2581 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2641 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2701 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2761 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     2821 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2881 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2941 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3001 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3061 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3121 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3181 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3241 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3301 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3361 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3421 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3481 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3541 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3601 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgatg aaaaagcctg
     3661 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
     3721 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
     3781 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
     3841 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg
     3901 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg
     3961 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg
     4021 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
     4081 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
     4141 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
     4201 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
     4261 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
     4321 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
     4381 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt
     4441 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg
     4501 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg
     4561 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac
     4621 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagcacgtgc
     4681 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4741 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4801 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4861 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4921 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4981 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5041 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5101 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5161 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5221 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5281 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5341 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5401 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5461 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5521 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5581 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5641 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5701 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5761 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5821 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5881 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5941 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6001 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6061 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6121 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6181 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6241 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6301 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6361 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6421 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6481 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6541 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6601 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6661 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6721 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6781 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6841 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6901 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6961 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7021 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7081 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     7141 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7201 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     7261 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     7321 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtt
//
