LOCUS       sequence-311021-        8125 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tagging clonal populations with whole cell mTAGBFP2 fluorescent 
            protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8125)
  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 8125)
  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: mTAGBFP2 WC visual barcode
FEATURES             Location/Qualifiers
     source          1..8125
                     /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             1251..1268
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1278..1979
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    1997..2585
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2050..2070)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2468..2479)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2657..2890
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2968..3089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3005..3024)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3059..3078
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3129..3264
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3284..3303)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3285..3303)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3313..3330)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3313..3329)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3322..3344)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3471..3926
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3558..3577)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3768..3789
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3952..4056
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4057..4125,4126..4917)
                     /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(4275..4294)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5088..5676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5577..5596
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5830..5847
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5964..5985
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6000..6005,6006..6023,6024..6030)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6038..6054
                     /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     6043..6065
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6062..6078
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6062..6078
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6097..6117
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6099..6117
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6145..6371
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6372..6552
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6599..6724
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7217..7450
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7635..7679
                     /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    7892..8009
                     /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 catggggagc caccatcacc
     1261 atcaccatgg cagatctatg agcgagctga ttaaggagaa catgcacatg aagctgtaca
     1321 tggagggcac cgtggacaac catcacttca agtgcacatc cgagggcgaa ggcaagccct
     1381 acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct
     1441 tcgacatcct ggctactagc ttcctctacg gcagcaagac cttcatcaac cacacccagg
     1501 gcatccccga cttcttcaag cagtccttcc ctgagggctt cacatgggag agagtcacca
     1561 catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc
     1621 tcatctacaa cgtcaagatc agaggggtga acttcacatc caacggccct gtgatgcaga
     1681 agaaaacact cggctgggag gccttcaccg agacgctgta ccccgctgac ggcggcctgg
     1741 aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag ccatctgatc gcaaacgcca
     1801 agaccacata tagatccaag aaacccgcta agaacctcaa gatgcctggc gtctactatg
     1861 tggactacag actggaaaga atcaaggagg ccaacaacga gacctacgtc gagcagcacg
     1921 aggtggcagt ggccagatac tgcgacctcc ctagcaaact ggggcacaag cttaattaaa
     1981 gacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2041 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2101 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2161 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2221 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2281 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2341 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     2401 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     2461 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     2521 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     2581 cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     2641 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     2701 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     2761 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     2821 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     2881 atctctagca gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca
     2941 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     3001 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3061 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     3121 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3181 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3241 agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta
     3301 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     3361 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     3421 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     3481 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3541 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3601 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3661 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3721 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3781 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3841 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3901 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     3961 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4021 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4081 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4141 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4201 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4261 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4321 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4381 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4441 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4501 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4561 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4621 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4681 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4741 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4801 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4861 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4921 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4981 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5041 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5101 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5161 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5221 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5281 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5341 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5401 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5461 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5521 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5581 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5641 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5701 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5761 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5821 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     5881 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     5941 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     6001 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     6061 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
     6121 caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca
     6181 tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat
     6241 tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat
     6301 ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc
     6361 gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     6421 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     6481 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     6541 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
     6601 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
     6661 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
     6721 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
     6781 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
     6841 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
     6901 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
     6961 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
     7021 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac
     7081 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     7141 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     7201 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     7261 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     7321 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     7381 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     7441 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     7501 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     7561 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     7621 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     7681 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     7741 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     7801 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga caattctaga
     7861 tcttgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt
     7921 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     7981 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccac
     8041 tttggcgccg gctcgagggg gcccggggtt attaatagta atcaattacg gggtcattag
     8101 ttcatagccc atatatggag ttccg
//
