LOCUS       sequence-414097-        7487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector for cloning a gene of interest with a 
            glycine-serine linker and TurboID on the N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7487)
  AUTHORS   Day RB, Hickman JA, Xu Z, Katerndahl CD, Ferraro F, Ramakrishnan SM,
            Erdmann-Gilmore P, Sprung RW, Mi Y, Townsend RR, Miller CA, Ley TJ
  TITLE     Proteogenomic analysis reveals cytoplasmic sequestration of RUNX1 by
            the acute myeloid leukemia-initiating CBFB::MYH11 oncofusion 
            protein.
  JOURNAL   J Clin Invest. 2023 Dec 7:e176311. doi: 10.1172/JCI176311.
  PUBMED    38061017
REFERENCE   2  (bases 1 to 7487)
  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: "J Clin 
            Invest. 2023 Dec 7:e176311. doi: 10.1172/JCI176311."
COMMENT     Sequence Label: TurboID-MCS MSCV-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..7487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    580..921
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             988..1404
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1334..1356
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1372..1388
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1441..2400
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     misc_feature    2496..3048
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2640..2657)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2867..2886
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3060..3062,3063..3065,3066..3779)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3366..3385)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3713..3734
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3881..4395
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4557..4573
                     /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(4581..4587,4588..4605,4606..4611))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4626..4647
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4764..4781)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4935..5523)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5015..5034)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5694..6485,6486..6554))
                     /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(6555..6659)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6727..6745
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6783..6805)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6905..6924
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7118..7140
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7268..7287
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7487,1..516)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
       61 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
      121 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
      181 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
      241 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      541 acccctgccc agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
      841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
      901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
      961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1381 gccctcactc cttctctagg cgccggaatt agatctctcg aggttaacca attcgccacc
     1441 atgaaagaca atactgtgcc tctgaagctg atcgctctcc tggctaatgg cgagttccat
     1501 agtggcgaac agctgggaga aaccctgggc atgtccaggg ccgctatcaa caagcacatt
     1561 cagactctgc gcgactgggg cgtggacgtg ttcaccgtgc ccggaaaggg ctactctctg
     1621 cccgagccta tcccgctgct gaacgctaaa cagattctgg gacagctgga cggcgggagc
     1681 gtggcagtcc tgcctgtggt cgactccacc aatcagtacc tgctggatcg aatcggcgag
     1741 ctgaagagtg gggatgcttg cattgcagaa tatcagcagg cagggagagg aagcagaggg
     1801 aggaaatggt tctctccttt tggagctaac ctgtacctga gtatgttttg gcgcctgaag
     1861 cggggaccag cagcaatcgg cctgggcccg gtcatcggaa ttgtcatggc agaagcgctg
     1921 cgaaagctgg gagcagacaa ggtgcgagtc aaatggccca atgacctgta tctgcaggat
     1981 agaaagctgg caggcatcct ggtggagctg gccggaataa caggcgatgc tgcacagatc
     2041 gtcattggcg ccgggattaa cgtggctatg aggcgcgtgg aggaaagcgt ggtcaatcag
     2101 ggctggatca cactgcagga agcagggatt aacctggaca ggaatactct ggccgctacg
     2161 ctgatccgag agctgcgggc agccctggaa ctgttcgagc aggaaggcct ggctccatat
     2221 ctgccacggt gggagaagct ggataacttc atcaatagac ccgtgaagct gatcattggg
     2281 gacaaagaga ttttcgggat tagccggggg attgataaac agggagccct gctgctggaa
     2341 caggacggag ttatcaaacc ctggatgggc ggagaaatca gtctgcggtc tgccgaaaag
     2401 ggtggaggtg gttctggcgg aggcggctct ggaggaggag gaagtggatc ctcgcgaacc
     2461 ggtatgcatc cgcgggaatt cgcccccccc ccctaacgtt actggccgaa gccgcttgga
     2521 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     2581 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     2641 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     2701 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     2761 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     2821 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     2881 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     2941 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     3001 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca
     3061 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     3121 gcgacgtaaa cggccacaag ttcagcgtgt ctggcgaggg cgagggcgat gccacctacg
     3181 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     3241 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     3301 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3361 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3421 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3481 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     3541 gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     3601 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3661 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3721 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat
     3781 gaattaatta agaattatca agcttatcga taccgtcgac ctgcagccaa gcttatcgat
     3841 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     3901 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga
     3961 atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga
     4021 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     4081 ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     4141 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     4201 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
     4261 ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     4321 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     4381 gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg
     4441 agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat
     4501 actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt gtgaaattgt
     4561 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4621 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4681 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4741 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4801 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4861 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4921 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4981 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5041 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5101 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5161 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5221 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5281 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5341 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5401 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5461 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5521 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5581 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5641 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5701 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5761 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5821 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5881 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5941 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6001 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6061 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6121 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6181 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6241 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6301 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6361 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6421 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6481 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6541 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6601 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6661 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     6721 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     6781 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     6841 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     6901 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     6961 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     7021 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     7081 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     7141 acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     7201 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     7261 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     7321 cggccacgat gcgtccggcg tagaggcgat tagtccaatt tgttaaagac aggatatcag
     7381 tggtccaggc tctagttttg actcaacaat atcaccagct gaagcctata gagtacgagc
     7441 catagataaa ataaaagatt ttatttagtc tccagaaaaa gggggga
//
