LOCUS       sequence-300189-        5585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses split-TuboID N-terminal fragment fused to GCGR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5585)
  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 5585)
  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: pAAV CMV GCGR-V5-sTurboID (N)
FEATURES             Location/Qualifiers
     source          1..5585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        201..504
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        505..708
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     659..679
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     753..772
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        753..771
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2276..2317
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2606..2623)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2612..2819
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2848..2988
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2848..2977
                     /label=AAV2 ITR
     rep_origin      3063..3518
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3150..3169)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3360..3381
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3535..3554)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3654..3676
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3714..3732)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3800..3904
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3905..3973,3974..4765)
                     /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(4123..4142)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4936..5524
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5425..5444
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaca tagtaatcaa ttacggggtc attagttcat
      181 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      241 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      301 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      361 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      421 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      481 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      541 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      601 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      661 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
      721 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
      781 gcttgccacc atgcccccct gccagccaca gcgacccctg ctgctgttgc tgctgctgct
      841 ggcctgccag ccacaggtcc cctccgctca ggtgatggac ttcctgtttg agaagtggaa
      901 gctctacggt gaccagtgtc accacaacct gagcctgctg ccccctccca cggagctggt
      961 gtgcaacaga accttcgaca agtattcctg ctggccggac acccccgcca ataccacggc
     1021 caacatctcc tgcccctggt acctgccttg gcaccacaaa gtgcaacacc gcttcgtgtt
     1081 caagagatgc gggcccgacg gtcagtgggt gcgtggaccc cgggggcagc cttggcgtga
     1141 tgcctcccag tgccagatgg atggcgagga gattgaggtc cagaaggagg tggccaagat
     1201 gtacagcagc ttccaggtga tgtacacagt gggctacagc ctgtccctgg gggccctgct
     1261 cctcgccttg gccatcctgg ggggcctcag caagctgcac tgcacccgca atgccatcca
     1321 cgcgaatctg tttgcgtcct tcgtgctgaa agccagctcc gtgctggtca ttgatgggct
     1381 gctcaggacc cgctacagcc agaaaattgg cgacgacctc agtgtcagca cctggctcag
     1441 tgatggagcg gtggctggct gccgtgtggc cgcggtgttc atgcaatatg gcatcgtggc
     1501 caactactgc tggctgctgg tggagggcct gtacctgcac aacctgctgg gcctggccac
     1561 cctccccgag aggagcttct tcagcctcta cctgggcatc ggctggggtg cccccatgct
     1621 gttcgtcgtc ccctgggcag tggtcaagtg tctgttcgag aacgtccagt gctggaccag
     1681 caatgacaac atgggcttct ggtggatctt gcggttcccc gtcttcctgg ccatcctgat
     1741 caacttcttc atcttcgtcc gcatcgttca gctgctcgtg gccaagctgc gggcacggca
     1801 gatgcaccac acagactaca agttccggct ggccaagtcc acgctgaccc tcatccctct
     1861 gctgggcgtc cacgaagtgg tctttgcctt cgtgacggac gagcacgccc agggcaccct
     1921 gcgctccgcc aagctcttct tcgacctctt cctcagctcc ttccagggcc tgctggtggc
     1981 tgtcctctac tgcttcctca acaaggaggt gcagtcggag ctgcggcggc gttggcaccg
     2041 ctggcgcctg ggcaaagtgc tatgggagga gcggaacacc agcaaccaca gggcctcatc
     2101 ttcgcccggc cacggccctc ccagcaagga gctgcagttt gggaggggtg gtggcagcca
     2161 ggattcatct gcggagaccc ccttggctgg tggcctccct agattggctg agagcccctt
     2221 cactagtgag ctggccgaga agctggccgg cctggacatc aacggtggag ctagcggcaa
     2281 gcccatcccc aaccccctgc tgggcctgga cagcaccaag ggctcgggct cgacctcggg
     2341 ctcgggcacc ggtaaagaca atactgtgcc tctgaagctg atcgctctcc tggctaatgg
     2401 cgagttccat agtggcgaac agctgggaga aaccctgggc atgtccaggg ccgctatcaa
     2461 caagcacatt cagactctgc gcgactgggg cgtggacgtg ttcaccgtgc ccggaaaggg
     2521 ctactctctg cccgagccta tcccgctgct gaacgctaaa cagattctgt aataagaatt
     2581 cctcgagcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2641 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2701 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2761 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggag
     2821 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     2881 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     2941 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     3001 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     3061 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3121 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3181 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3241 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     3301 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3361 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     3421 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     3481 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     3541 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3601 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3661 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3721 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3781 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3841 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3901 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3961 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4021 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4081 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4141 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4201 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4261 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4321 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4381 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4441 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4501 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4561 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4621 aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4681 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4741 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4801 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4861 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4921 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4981 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5041 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5101 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5161 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5221 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5281 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5341 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5401 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5461 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5521 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5581 catgt
//
