LOCUS       sequence-311030-        8116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tagging clonal populations with Peroxisomal mTurquoise2 fluorescent 
            protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8116)
  AUTHORS   Kaufman T, Nitzan E, Firestein N, Ginzberg MB, Iyengar S, Patel N, 
            Ben-Hamo R, Porat Z, Hunter J, Hilfinger A, Rotter V, Kafri R, 
            Straussman R
  TITLE     Visual barcodes for clonal-multiplexing of live microscopy-based 
            assays.
  JOURNAL   Nat Commun. 2022 May 18;13(1):2725. doi: 10.1038/s41467-022-30008-0.
  PUBMED    35585055
REFERENCE   2  (bases 1 to 8116)
  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: "Nat 
            Commun."; date: "2022-05-18"; volume: "13(1):2725. doi"; pages: " 
            10.1038/s41467-022-30008-0"
COMMENT     Sequence Label: mTurquoise2 Peroxisome visual barcode
FEATURES             Location/Qualifiers
     source          1..8116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(534..553)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1030..1050
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1170..1235
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             join(1242..1244,1245..1247,1248..1958)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1287..1308)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1548..1567)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1895..1916
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1988..2576
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2041..2061)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2459..2470)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2648..2881
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2959..3080
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2996..3015)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3050..3069
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3120..3255
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3275..3294)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3276..3294)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3304..3321)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3304..3320)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3313..3335)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3462..3917
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3549..3568)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3759..3780
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3943..4047
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4048..4116,4117..4908)
                     /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(4266..4285)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5079..5667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5568..5587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5821..5838
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5955..5976
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5991..5996,5997..6014,6015..6021)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6029..6045
                     /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     6034..6056
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6053..6069
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6053..6069
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6088..6108
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6090..6108
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6136..6362
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6363..6543
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6590..6715
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7208..7441
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7626..7670
                     /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"
     misc_feature    7883..8000
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ctggctaact gtcgggatcc accatgaccg
      541 agtacaagcc caccgtgcgc ctcgccaccc gcgacgacgt gcccagggcc gtccgcaccc
      601 tcgccgccgc tttcgccgac taccccgcca cccgccacac cgtcgatccc gaccgccaca
      661 tcgagcgggt caccgagctg caagaactct tcctcacccg cgtcgggctc gacatcggca
      721 aggtgtgggt cgctgacgac ggcgccgcag tggctgtctg gaccacaccc gagagcgtcg
      781 aagctggggc ggtgttcgcc gagatcggcc ctcgcatggc cgagttgagc ggttcccggc
      841 tggccgctca gcaacagatg gaaggcctcc tggctccaca ccggcccaag gagcccgcat
      901 ggttcctggc caccgtcggc gtctctcccg accaccaggg caagggtctg ggcagcgccg
      961 tcgtgctccc cggagtggag gctgccgagc gcgccggggt gcccgccttc ctggagacct
     1021 ccgcaccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtgg
     1081 aggtgcccga aggaccacgc acctggtgca tgacccgcaa gcccggtgcc ctgcagggat
     1141 ctggcgaagc cagacacaaa cagaaaattg tgaaacagac tttgaatttt gaccttctca
     1201 agttggcggg agacgtggag tccaacccag ggcccgctag catggtgagc aagggcgagg
     1261 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1321 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1381 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgtcct
     1441 ggggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1501 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1561 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1621 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactact
     1681 ttagcgacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     1741 agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca
     1801 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca
     1861 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1921 ccgccgggat cactctcggc atggacgagc tgtacaagtc ccgcctctaa agacgcgtta
     1981 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2041 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2101 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2161 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2221 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2281 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2341 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     2401 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     2461 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     2521 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg
     2581 actttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     2641 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     2701 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     2761 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     2821 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     2881 agtacgtata gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     2941 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3001 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3061 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     3121 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3181 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3241 gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg
     3301 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     3361 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     3421 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     3481 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3541 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3601 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3661 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3721 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3781 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3841 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3901 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3961 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4021 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4081 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4141 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4201 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4261 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4321 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4381 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4441 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4501 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4561 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4621 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4681 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4741 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4801 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4861 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4921 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4981 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5041 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5101 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5161 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5221 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5281 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5341 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5401 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5461 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5521 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5581 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5641 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5701 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5761 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5821 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     5881 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     5941 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     6001 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     6061 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg
     6121 gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga
     6181 tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag
     6241 taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac
     6301 gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa
     6361 acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     6421 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     6481 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     6541 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
     6601 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
     6661 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
     6721 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     6781 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     6841 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     6901 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     6961 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     7021 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
     7081 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     7141 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     7201 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     7261 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     7321 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     7381 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     7441 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     7501 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     7561 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     7621 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     7681 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     7741 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     7801 caccattatc gtttcagacc cacctcccaa ccccgagggg acaattctag atcttgagac
     7861 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     7921 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     7981 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca ctttggcgcc
     8041 ggctcgaggg ggcccggggt tattaatagt aatcaattac ggggtcatta gttcatagcc
     8101 catatatgga gttccg
//
