LOCUS       sequence-232589-        7419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ePiggyBac mammalian expression plasmid for tandem dimer 
            MS2cp-tagRFP-T fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7419)
  AUTHORS   Alexander JM, Guan J, Li B, Maliskova L, Song M, Shen Y, Huang B, 
            Lomvardas S, Weiner OD
  TITLE     Live-cell imaging reveals enhancer-dependent Sox2 transcription in 
            the absence of enhancer proximity.
  JOURNAL   Elife. 2019 May 24;8. pii: 41769. doi: 10.7554/eLife.41769.
  PUBMED    31124784
REFERENCE   2  (bases 1 to 7419)
  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: "Elife."; 
            date: "2019-05-24"; volume: "8. pii: 41769. doi"; pages: " 
            10.7554/eLife.41769"
COMMENT     Sequence Label: epB_UbC_tdMS2cp-tagRFP-Tx2
FEATURES             Location/Qualifiers
     source          1..7419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        103..1314
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1151..1170
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    1331..1355
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1425..1451
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             1425..1451
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1452..1463
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2232..2963
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     CDS             2970..3701
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     CDS             3702..3722
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(3757..3781)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3897..4018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3934..3953)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3988..4007
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4204..4266
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4301..4320)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4302..4320)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4330..4347)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4330..4346)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4339..4361)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4488..4943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4575..4594)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4785..4806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4969..5073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5074..5142,5143..5934)
                     /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(5292..5311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6105..6693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6594..6613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6847..6864
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6981..7002
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7017..7022,7023..7040,7041..7047)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7055..7071
                     /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     7060..7082
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7079..7095
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7079..7095
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7114..7134
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7116..7134
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   7182..7216
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 aattatattg ttatttcgtg ttctacttac gtgataactt attatatata tattttcttg
       61 ttatagatat ccttctcgac attgattatt gactagagat ctggcctccg cgccgggttt
      121 tggcgcctcc cgcgggcgcc cccctcctca cggcgagcgc tgccacgtca gacgaagggc
      181 gcagcgagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag
      241 actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact
      301 ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg
      361 gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc
      421 cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg
      481 atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc
      541 cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg
      601 tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt
      661 tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg
      721 gggggcatgg tgggcggcaa gaacccaagg tcttgaggcc ttcgctaatg cgggaaagct
      781 cttattcggg tgagatgggc tggggcacca tctggggacc ctgacgtgaa gtttgtcact
      841 gactggagaa ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg
      901 cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg
      961 ttggcttata atgcagggtg gggccacctg ccggtaggtg tgcggtaggc ttttctccgt
     1021 cgcaggacgc agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc
     1081 tggtgagggg agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt
     1141 cttaagtagc tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt
     1201 gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt
     1261 tagactagta aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacgaagct
     1321 cgaggatatc acaagtttgt acaaaaaagc aggctttaaa ggaaccaatt cagtcgactg
     1381 gatccggtac cgaattcgcg gccgcctagc taaccaccat gggctacccc tacgacgtgc
     1441 ccgactacgc catcgaaggc cgccatatgc tagccgttaa aatggcttct aactttactc
     1501 agttcgttct cgtcgacaat ggcggaactg gcgacgtgac tgtcgcccca agcaacttcg
     1561 ctaacgggat cgctgaatgg atcagctcta actcgcgttc acaggcttac aaagtaacct
     1621 gtagcgttcg tcagagctct gcgcagaatc gcaaatacac catcaaagtc gaggtgccta
     1681 aaggcgcctg gcgttcgtac ttaaatatgg aactaaccat tccaattttc gccacgaatt
     1741 ccgactgcga gcttattgtt aaggcaatgc aaggtctcct aaaagatgga aacccgattc
     1801 cctcagcaat cgcagcaaac tccggcatct acgcgatcta cgccatggct tctatggctt
     1861 ctaactttac tcagttcgtt ctcgtcgaca atggcggaac tggcgacgtg actgtcgccc
     1921 caagcaactt cgctaacggg atcgctgaat ggatcagctc taactcgcgt tcacaggctt
     1981 acaaagtaac ctgtagcgtt cgtcagagct ctgcgcagaa tcgcaaatac accatcaaag
     2041 tcgaggtgcc taaaggcgcc tggcgttcgt acttaaatat ggaactaacc attccaattt
     2101 tcgccacgaa ttccgactgc gagcttattg ttaaggcaat gcaaggtctc ctaaaagatg
     2161 gaaacccgat tccctcagca atcgcagcaa actccggcat ctacgcggat ccaccggtcg
     2221 ccaccggatc tatggtgtct aagggcgaag agctgattaa ggagaacatg cacatgaagc
     2281 tgtacatgga gggcaccgtg aacaaccacc acttcaagtg cacatccgag ggcgaaggca
     2341 agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct
     2401 tcgccttcga catcctggct accagcttca tgtacggcag cagaaccttc atcaaccaca
     2461 cccagggcat ccccgatttc tttaagcagt ccttccctga gggcttcaca tgggagagag
     2521 tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg
     2581 gctgcctcat ctacaacgtc aagatcagag gggtgaactt cccatccaac ggccctgtga
     2641 tgcagaagaa aacactcggc tgggaggcca acaccgagat gctgtacccc gctgacggcg
     2701 gcctggaagg cagaaccgac atggccctga agctcgtggg cgggggccac ctgatctgca
     2761 acttcaagac cacatacaga tccaagaaac ccgctaagaa cctcaagatg cccggcgtct
     2821 actatgtgga ccacagactg gaaagaatca aggaggccga caaagagacg tacgtcgagc
     2881 agcacgaggt ggctgtggcc agatactgcg acctccctag caaactgggg cacaaactta
     2941 atggcatgga cgagctgtac aagctaggga tggtgtctaa gggcgaagag ctgattaagg
     3001 agaacatgca catgaagctg tacatggagg gcaccgtgaa caaccaccac ttcaagtgca
     3061 catccgaggg cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg
     3121 agggcggccc tctccccttc gccttcgaca tcctggctac cagcttcatg tacggcagca
     3181 gaaccttcat caaccacacc cagggcatcc ccgatttctt taagcagtcc ttccctgagg
     3241 gcttcacatg ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg
     3301 acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc
     3361 catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccaac accgagatgc
     3421 tgtaccccgc tgacggcggc ctggaaggca gaaccgacat ggccctgaag ctcgtgggcg
     3481 ggggccacct gatctgcaac ttcaagacca catacagatc caagaaaccc gctaagaacc
     3541 tcaagatgcc cggcgtctac tatgtggacc acagactgga aagaatcaag gaggccgaca
     3601 aagagacgta cgtcgagcag cacgaggtgg ctgtggccag atactgcgac ctccctagca
     3661 aactggggca caaacttaat ggcatggacg agctgtacaa gcctaagaaa aagcggaaag
     3721 tgtaagtgcc taggcggccg cactcgagat atctagaccc agctttcttg tacaaagtgg
     3781 tgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     3841 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     3901 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3961 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     4021 gaattcgata aaagttttgt tactttatag aagaaatttt gagtttttgt ttttttttaa
     4081 taaataaata aacataaata aattgtttgt tgaatttatt attagtatgt aagtgtaaat
     4141 ataataaaac ttaatatcta ttcaaattaa taaataaacc tcgatataca gaccgataaa
     4201 acacatgcgt caattttacg catgattatc tttaacgtac gtcacaatat gattatcttt
     4261 ctagggttaa tctagctgcg gcgcgccggt acccaattcg ccctatagtg agtcgtatta
     4321 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     4381 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     4441 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     4501 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4561 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4621 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4681 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4741 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4801 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4861 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4921 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     4981 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5041 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5101 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5161 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5221 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5281 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5341 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5401 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5461 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5521 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5581 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5641 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5701 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5761 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5821 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5881 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5941 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6001 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6061 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6121 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6181 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6241 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6301 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6361 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6421 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6481 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6541 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6601 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6661 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6721 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6781 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6841 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6901 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6961 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7021 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7081 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
     7141 aagctggagc tccaccgcgg ggcgcgccgc agctagatta accctagaaa gatagtctgc
     7201 gtaaaattga cgcatgcatt cttgaaatat tgctctctct ttctaaatag cgcgaatccg
     7261 tcgctgtgca tttaggacat ctcagtcgcc gcttggagct cccgtgaggc gtgcttgtca
     7321 atgcggtaag tgtcactgat tttgaactat aacaaccgcg tgagtcaaaa tgacgcatga
     7381 ttatctttta cgtgactttt aagatttaac tcatacgat
//
