LOCUS       sequence-358003-        8310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses low-affinity split-TurboID N-terminal fragment in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8310)
  AUTHORS   Cho KF, Branon TC, Rajeev S, Svinkina T, Udeshi ND, Thoudam T, Kwak 
            C, Rhee HW, Lee IK, Carr SA, Ting AY
  TITLE     Split-TurboID enables contact-dependent proximity labeling in cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 May 18. pii: 1919528117. doi: 
            10.1073/pnas.1919528117.
  PUBMED    32424107
REFERENCE   2  (bases 1 to 8310)
  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 18. pii: 1919528117. doi: 
            10.1073/pnas.1919528117."
COMMENT     Sequence Label: pLX304 CMV FKBP-V5-sTurboID (N)
FEATURES             Location/Qualifiers
     source          1..8310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     387..404
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    521..542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(557..562,563..580,581..587)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    595..611
                     /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     600..622
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     619..635
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     619..635
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     654..674
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        656..674
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        702..928
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             929..1109
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1156..1281
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1774..2007
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2192..2236
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        2411..2911
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2830..2848
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2923..3318
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    3378..3495
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3612..3915
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3916..4119
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4070..4090
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    4142..4166
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4532..4573
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    4864..5452
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4917..4937)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5335..5346)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5524..5757
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5835..5956
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5872..5891)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5926..5945
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5996..6131
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6151..6170)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6152..6170)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6180..6197)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6180..6196)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6189..6211)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6338..6793
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6425..6444)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6635..6656
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6819..6923
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6924..6992,6993..7784)
                     /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(7142..7161)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7955..8310,1..233)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
       61 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      121 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      181 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      241 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      301 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      361 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      421 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
      481 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
      541 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
      601 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
      661 accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt atgcaatact
      721 cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa
      781 gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca
      841 acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt
      901 atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga tctgagcctg
      961 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     1021 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     1081 cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa
     1141 gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga
     1201 ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag
     1261 agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa
     1321 attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc
     1381 agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga
     1441 caaatactgg gacagctaca accatccctt cagacaggat cagaagaact tagatcatta
     1501 tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag
     1561 gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg
     1621 gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata
     1681 taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag
     1741 agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg
     1801 agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt
     1861 attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca
     1921 tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga
     1981 aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg
     2041 caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa
     2101 tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct taatacactc
     2161 cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga
     2221 taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt
     2281 attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat
     2341 agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc
     2401 gaggggaccc ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc
     2461 tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc
     2521 cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc
     2581 tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac
     2641 ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc
     2701 gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag
     2761 cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct
     2821 gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct
     2881 cgttgaccga atcaccgacc tctctcccca gggggtacca ccatggccaa gcctttgtct
     2941 caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct
     3001 gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc
     3061 aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct
     3121 gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc
     3181 ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc
     3241 atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct
     3301 ggttatgtgt gggagggcct gcagctgcag tagtaagaat tctagatctt gagacaaatg
     3361 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     3421 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     3481 aaaattcaaa attttcgggt ttattacagg gacagcagag atccactttg gcgccggctc
     3541 gagggggccc ggggttatta atagtaatca attacggggt cattagttca tagcccatat
     3601 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     3661 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     3721 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     3781 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     3841 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     3901 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     3961 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     4021 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     4081 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tgtcgggatc
     4141 aacaagtttg tacaaaaaag caggcttcga aggagataga accatgcagg tggaaaccat
     4201 ctccccagga gacgggcgca ccttccccaa gcgcggccag acctgcgtgg tgcactacac
     4261 cgggatgctt gaagatggaa agaaatttga ttcctcccgg gacagaaaca agccctttaa
     4321 gtttatgcta ggcaagcagg aggtgatccg aggctgggaa gaaggggttg cccagatgag
     4381 tgtgggtcag agagccaaac tgactatatc tccagattat gcctatggtg ccactgggca
     4441 cccaggcatc atcccaccac atgccactct cgtcttcgat gtggagcttc taaaactgga
     4501 agggagtggc agcggaggta gtgggagcgg tggcaagccc atccccaacc ccctgctggg
     4561 cctggacagc accaagggct cgggctcgac ctcgggctcg ggcaccggta aagacaatac
     4621 tgtgcctctg aagctgatcg ctctcctggc taatggcgag ttccatagtg gcgaacagct
     4681 gggagaaacc ctgggcatgt ccagggccgc tatcaacaag cacattcaga ctctgcgcga
     4741 ctggggcgtg gacgtgttca ccgtgcccgg aaagggctac tctctgcccg agcctatccc
     4801 gctgctgaac gctaaacaga ttctgtaata agctagcgct aaccggtggc gcgttaagtc
     4861 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     4921 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     4981 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     5041 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     5101 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     5161 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     5221 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     5281 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     5341 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     5401 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcgtcgactt
     5461 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     5521 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     5581 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     5641 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     5701 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     5761 cgtatagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     5821 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5881 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5941 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc
     6001 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6061 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6121 ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     6181 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     6241 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     6301 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     6361 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     6421 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     6481 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     6541 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     6601 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     6661 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     6721 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     6781 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     6841 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     6901 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     6961 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     7021 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     7081 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     7141 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     7201 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     7261 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     7321 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     7381 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     7441 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     7501 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     7561 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     7621 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     7681 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     7741 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     7801 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     7861 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7921 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     7981 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     8041 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     8101 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     8161 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     8221 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     8281 gttcgtgcac acagcccagc ttggagcgaa
//
