LOCUS       sequence-369088-        8354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RNAP2 Ser5ph-mintbody expression vector for mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8354)
  AUTHORS   Ohishi H, Shimada S, Uchino S, Li J, Sato Y, Shintani M, Owada H, 
            Ohkawa Y, Pertsinidis A, Yamamoto T, Kimura H, Ochiai H
  TITLE     STREAMING-tag system reveals spatiotemporal relationships between 
            transcriptional regulatory factors and transcriptional activity.
  JOURNAL   Nat Commun. 2022 Dec 20;13(1):7672. doi: 10.1038/s41467-022-35286-2.
  PUBMED    36539402
REFERENCE   2  (bases 1 to 8354)
  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: "2022-12-20"; volume: "13(1):7672. doi"; pages: " 
            10.1038/s41467-022-35286-2"
COMMENT     Sequence Label: PB533-44B12m23-sfGFP
FEATURES             Location/Qualifiers
     source          1..8354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1389..1600
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1613..1881
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      2674..2683
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2680..2682,2683..2685,2686..3399)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2725..2746)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3333..3354
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(3409..3983,3984..3986,3987..3995)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3576..3593)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3803..3822
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4008..4802
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLAATGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4062..4081)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4672..4691
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4862..4882)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4924..5045
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4961..4980)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5015..5034
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5808..5842
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6333..6349)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6333..6349)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6346..6368)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6357..6373
                     /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(6381..6387,6388..6405,6406..6411))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6426..6447
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6564..6581)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6735..7323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6815..6834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7494..8285,8286..8354))
                     /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     8117..8136
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccaagg atctgcgatc gctccggtgc
     1381 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     1441 caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     1501 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     1561 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg
     1621 catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg
     1681 cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt
     1741 aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag
     1801 ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc
     1861 tgttctgcgc cgttacagat ccaagctgtg accggcgcct actctagagc tagcgaattc
     1921 gccatggccg aagtccagct gcaacagtct ggaccggagc tggtgaagcc tggggcttca
     1981 gtgaagatat cctgcaaggc ttctggatac acattcactg aatacaccat gcactgggtg
     2041 aaacagagcc atggacagag ccttgagtgg attggaggca ttaatcctaa caatgttctt
     2101 acttactaca accagaggtt caagggcaag gccacattga ctgtagacaa gtcctccagc
     2161 acagcctacc tggagctccg cagcctgaca tctgaggatt ctgcagtcta ttactgtgca
     2221 agaggcacct cttttgctta ctggggccaa gggactctgg tcactgtctc ctcaggtgga
     2281 ggcggttcag gcggaggtgg ctctggcggt ggcggatcgg atgttgtgat gacccagact
     2341 ccactcactt tgtcggttac cattggacaa ccagcctcca tctcttgcaa gtcaagtcag
     2401 agcctcttat atagtgatgg aaagacatat ttgagttggt tgttacagag gccaggccag
     2461 tctccagagc gcctaatcta tctggtgtct aaactggact ctggagtccc tgacaggttc
     2521 actggcagtg gatcagggac agatttcaca ctgaaaatca gcagagtgga ggctgaggat
     2581 ttgggagttt attattgctg gcaaggtaca cattttcctc ggacgttcgg tggaggcacc
     2641 aagctggaaa taaaacgggc ggatccaccg gtcgccacca tggtgagcaa gggcgaggag
     2701 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2761 ttcagcgtgc gcggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc
     2821 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     2881 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc gccacgactt cttcaagtcc
     2941 gccatgcccg aaggctacgt ccaggagcgc accatcagct tcaaggacga cggcacctac
     3001 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     3061 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac
     3121 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     3181 atccgccaca acgtggagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     3241 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgtg
     3301 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     3361 gccgggatca ctcacggcat ggacgagctg tacaagtaag cggccgcgcc cctctccctc
     3421 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     3481 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     3541 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     3601 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     3661 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     3721 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     3781 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     3841 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     3901 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     3961 cctttgaaaa acacgatgat aatatggcca caaccatggc gtccggaatg attgaacaag
     4021 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4081 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4141 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4201 cgcggctatc gtggctggcc gcgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4261 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4321 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4381 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4441 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4501 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     4561 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4621 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4681 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4741 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4801 gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     4861 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgcgttaa
     4921 ctaaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4981 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5041 tcttatcatg tctggaattg actcaaatga tgtcaattag tctatcagaa gctcatctgg
     5101 tctcccttcc gggggacaag acatccctgt ttaatattta aacagcagtg ttcccaaact
     5161 gggttcttat atcccttgct ctggtcaacc aggttgcagg gtttcctgtc ctcacaggaa
     5221 cgaagtccct aaagaaacag tggcagccag gtttagcccc ggaattgact ggattccttt
     5281 tttagggccc attggtatgg ctttttcccc gtatcccccc aggtgtctgc aggctcaaag
     5341 agcagcgaga agcgttcaga ggaaagcgat cccgtgccac cttccccgtg cccgggctgt
     5401 ccccgcacgc tgccggctcg gggatgcggg gggagcgccg gaccggagcg gagccccggg
     5461 cggctcgctg ctgcccccta gcgggggagg gacgtaatta catccctggg ggctttgggg
     5521 gggggctgtc cctgatatct ataacaagaa aatatatata taataagtta tcacgtaagt
     5581 agaacatgaa ataacaatat aattatcgta tgagttaaat cttaaaagtc acgtaaaaga
     5641 taatcatgcg tcattttgac tcacgcggtc gttatagttc aaaatcagtg acacttaccg
     5701 cattgacaag cacgcctcac gggagctcca agcggcgact gagatgtcct aaatgcacag
     5761 cgacggattc gcgctattta gaaagagaga gcaatatttc aagaatgcat gcgtcaattt
     5821 tacgcagact atctttctag ggttaatcta gctgcatcag gatcatatcg tcgggtcttt
     5881 tttccggctc agtcatcgcc caagctggcg ctatctgggc atcggggagg aagaagcccg
     5941 tgccttttcc cgcgaggttg aagcggcatg gaaagagttt gccgaggatg actgctgctg
     6001 cattgacgtt gagcgaaaac gcacgtttac catgatgatt cgggaaggtg tggccatgca
     6061 cgcctttaac ggtgaactgt tcgttcaggc cacctgggat accagttcgt cgcggctttt
     6121 ccggacacag ttccggatgg tcagcccgaa gcgcatcagc aacccgaaca ataccggcga
     6181 cagccggaac tgccgtgccg gtgtgcagat taatgacagc ggtgcggcgc tgggatatta
     6241 cgtcagcgag gacgggtatc ctggctggat gccgcagaaa tggacatgga taccccgtga
     6301 gttacccggc gggcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6361 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6421 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6481 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6541 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6601 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6661 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6721 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6781 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6841 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6901 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6961 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7021 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7081 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7141 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7201 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7261 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7321 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7381 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7441 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7501 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7561 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7621 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7681 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7741 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7801 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7861 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7921 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7981 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8041 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8101 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8161 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8221 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8281 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8341 tgttgaatac tcat
//
