LOCUS       sequence-300192-        8137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses split-TurboID C-terminal fragment fused to arrestin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8137)
  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 8137)
  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: pCDNA3 CMV sTurboID (C)-Arrestin-GFP
FEATURES             Location/Qualifiers
     source          1..8137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2940..2942,2943..2945,2946..3659)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2940..2942,2943..2945,2946..3656)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3246..3265)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3593..3614
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3690..3708)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3691..3708)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3728..3745)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3734..3958
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4004..4432
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4091..4110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4301..4322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4441..4461)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4446..4775
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4626..4761
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4688..4707
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4842..5636
                     /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(4896..4915)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5506..5525
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5810..5931
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5847..5866)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5901..5920
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5980..5996)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5980..5996)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5993..6015)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6004..6020
                     /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(6028..6034,6035..6052,6053..6058))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6073..6094
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6211..6228)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6382..6970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6462..6481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7141..7932,7933..8001))
                     /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"
     promoter        complement(8002..8106)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgcggcc
      901 gcatgggaca gctggacggc gggagcgtgg cagtcctgcc tgtggtcgac tccaccaatc
      961 agtacctgct ggatcgaatc ggcgagctga agagtgggga tgcttgcatt gcagaatatc
     1021 agcaggcagg gagaggaagc agagggagga aatggttctc tccttttgga gctaacctgt
     1081 acctgagtat gttttggcgc ctgaagcggg gaccagcagc aatcggcctg ggcccggtca
     1141 tcggaattgt catggcagaa gcgctgcgaa agctgggagc agacaaggtg cgagtcaaat
     1201 ggcccaatga cctgtatctg caggatagaa agctggcagg catcctggtg gagctggccg
     1261 gaataacagg cgatgctgca cagatcgtca ttggcgccgg gattaacgtg gctatgaggc
     1321 gcgtggagga aagcgtggtc aatcagggct ggatcacact gcaggaagca gggattaacc
     1381 tggacaggaa tactctggcc gctacgctga tccgagagct gcgggcagcc ctggaactgt
     1441 tcgagcagga aggcctggct ccatatctgc cacggtggga gaagctggat aacttcatca
     1501 atagacccgt gaagctgatc attggggaca aagagatttt cgggattagc cgggggattg
     1561 ataaacaggg agccctgctg ctggaacagg acggagttat caaaccctgg atgggcggag
     1621 aaatcagtct gcggtctgcc gaaaagggct cgggcgggag taccggtggt agtgggagcg
     1681 gtggtgaaaa acccgggacc agggtcttca agaagtcgag ccctaactgc aagctcaccg
     1741 tgtacttggg caagcgtgac tttgtggatc acttggacaa agtggatcct gtcgatggtg
     1801 tggtgcttgt ggatcctgac tacttgaagg accggaaagt gtttgtgacc ctcacctgtg
     1861 ccttccgcta tggccgagaa gacctggatg tactgggcct gtctttccgc aaagatctgt
     1921 tcatcgccac ctaccaggcc ttccccccca tgcccaaccc acctcggccc cccacccgcc
     1981 tacaggaccg actgctgaag aagttgggcc agcacgccca cccctttttt ttcacaatac
     2041 cccagaattt gccttgctcc gtcacactgc agccaggacc ggaggacaca gggaaggcct
     2101 gtggagtaga ctttgagatt cgagccttct gtgccaaatc tatagaagaa aaaagccaca
     2161 aaaggaactc cgtgcggctt atcatcagaa aggtacagtt tgctcctgag acacccggcc
     2221 cccagccatc agctgaaacc acacgccact tcctcatgtc tgaccggagg tccctgcacc
     2281 tagaggcttc cctggacaaa gagctgtact accatgggga acccctcaat gtcaacgtcc
     2341 acgtcaccaa caattctgcc aaaaccgtca agaagatcag agtgtctgtg agacagtatg
     2401 ccgacatttg cctcttcagc accgcgcagt acaagtgtcc tgtggctcag cttgaacaag
     2461 atgaccaggt gtctcccagt tccacattct gcaaggtgta caccataacc ccgctgctca
     2521 gtgacaaccg agagaagcgt ggccttgccc ttgatgggca actcaagcac gaagacacca
     2581 acctggcttc cagcaccatt gtgaaggagg gagccaacaa ggaggtgctg ggaatcctag
     2641 tatcctacag ggtcaaggtg aagctggtgg tgtctcgagg cggggatgtc tccgtggagc
     2701 tacctttcgt cctaatgcac cccaagcccc acgaccacat cacccttccc cgaccccagt
     2761 cagccccccg ggaaatagac atccctgtgg ataccaacct cattgaattc gataccaact
     2821 atgccacaga cgacgacatc gtgtttgagg actttgcgag gcttcggctg aaggggatga
     2881 aggatgacga ctgtgatgac cagttctgcg gtggaagtgg atcaggcagc ggtggatcta
     2941 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     3001 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     3061 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     3121 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     3181 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3241 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3301 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3361 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     3421 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     3481 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3541 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3601 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat
     3661 aactcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc
     3721 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3781 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3841 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3901 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3961 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     4021 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4081 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4141 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4201 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4261 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4321 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     4381 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     4441 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4501 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4561 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4621 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4681 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4741 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4801 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4861 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4921 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4981 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     5041 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5101 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5161 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5221 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5281 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5341 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     5401 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5461 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5521 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5581 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5641 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5701 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5761 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5821 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5881 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5941 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6001 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6061 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6121 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6181 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6241 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6301 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6361 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6421 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6481 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6541 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6601 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac aaaccccccg ttcagcccga
     6661 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6721 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6781 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6841 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6901 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6961 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7021 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7081 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7141 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7201 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7261 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7321 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7381 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7441 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7501 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7561 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7621 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7681 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7741 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7801 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7861 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7921 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7981 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8041 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8101 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
