LOCUS       sequence-318470-        7037 bp    DNA     circular SYN 14-APR-2025
DEFINITION  GAVPO (VIVID transactivator) expressed under the CMV promoter in the
            pcDNA3.1 backbone. AKA 133.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7037)
  AUTHORS   Rodriques SG, Chen LM, Liu S, Zhong ED, Scherrer JR, Boyden ES, Chen
            F
  TITLE     RNA timestamps identify the age of single molecules in RNA 
            sequencing.
  JOURNAL   Nat Biotechnol. 2020 Oct 19. pii: 10.1038/s41587-020-0704-z. doi: 
            10.1038/s41587-020-0704-z.
  PUBMED    33077959
REFERENCE   2  (bases 1 to 7037)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2020 Oct 19. pii: 10.1038/s41587-020-0704-z. doi: 
            10.1038/s41587-020-0704-z."
COMMENT     Sequence Label: pcDNA3.1-GAVPO
FEATURES             Location/Qualifiers
     source          1..7037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1377..1403
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1833..2204
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     CDS             2205..2228
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2301..2342
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2352..2369
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2392..2409)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2398..2622
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2668..3096
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2755..2774)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2965..2986
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3105..3125)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3110..3439
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3290..3425
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3352..3371
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3506..4300
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3560..3579)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4170..4189
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4476..4597
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4513..4532)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4567..4586
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4646..4662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4646..4662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4659..4681)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4670..4686
                     /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(4694..4700,4701..4718,4719..4724))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4739..4760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4877..4894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5048..5636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5128..5147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5807..6598,6599..6667))
                     /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     6430..6449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6668..6772)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6847..6866)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag cttggtaccg agctcggatc cagtaccctt caccatgaag ttgttgagct
      721 caatcgaaca agcgtgtgac atatgtcgcc ttaagaaact caaatgttct aaagaaaagc
      781 ctaagtgcgc gaaatgtttg aaaaacaatt gggaatgccg ctattctcct aagacaaagc
      841 gctctccgct cacaagagcg caccttacag aggttgagtc ccgcctcgag cgcttggaga
      901 ggtccattgc aacgagaagt catacactct acgcccccgg agggtatgac ataatgggtt
      961 acctcatcca aataatgaaa agaccgaatc ctcaagtaga gcttggaccg gtcgacacat
     1021 ccgtagcact catactttgc gatttgaaac aaaaggacac tccgatagtg tatgcgtcag
     1081 aggcgtttct ttatatgaca gggtactcca acgccgaggt gttgggacga aattgccgat
     1141 ttcttcagtc cccagatgga atggttaagc ccaagtctac tagaaagtat gttgacagca
     1201 atacgattaa cactatgaga aaggccattg atcggaatgc cgaggtgcag gttgaggtag
     1261 tcaacttcaa gaagaatggg caaagattcg tcaacttcct tacaatgatt cccgttcgcg
     1321 atgagacggg cgagtatcga tactcaatgg gtttccagtg tgaaaccgaa ttgcaatatc
     1381 cttatgatgt acccgattac gcagaattcc aataccttcc agacacggat gatcgccatc
     1441 gaatcgaaga gaagagaaaa cgcacctacg agacgttcaa atctattatg aagaaatctc
     1501 ccttcagtgg gccgacggac ccaaggccgc caccgcgaag gatagccgtt ccatcaagaa
     1561 gctcagcttc tgtacccaaa cccgccccac agccttaccc ctttacttcc tccctctcca
     1621 ctatcaacta cgatgagttc cccacgatgg tttttccttc aggacagata tcccaagcga
     1681 gcgccctcgc accagcccca ccacaagtgc ttcctcaggc ccctgcgcct gctccggcac
     1741 cggcgatggt gagcgctctg gctcaagctc ccgcgccagt ccctgttttg gcaccagggc
     1801 cacctcaggc agtggctccg cccgctccaa aacctactca agcgggcgaa ggaactctga
     1861 gcgaggcgct cctgcagctt caatttgatg acgaagatct cggcgcactc ctcggtaatt
     1921 caacggaccc cgctgttttc actgacctgg caagcgtgga taactctgaa ttccaacagc
     1981 tccttaacca gggcataccg gtcgcgcctc atacaactga accaatgctg atggaatatc
     2041 cggaggcaat aaccagactt gtgacggggg cgcagcgacc gcctgatcca gcacccgcac
     2101 cgcttggggc gcctggcttg cccaatggac tcctttctgg cgacgaggac ttttccagca
     2161 tcgcagacat ggacttttct gcactccttt ctcagatttc ctcagattac aaagacgacg
     2221 atgataagaa gggtcaagac aattctgcag atatccagca cagtggcggc cgctcgagtc
     2281 tagagggccc gcggttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta
     2341 cgcgtaccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg
     2401 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2461 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2521 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2581 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2641 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2701 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2761 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2821 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2881 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2941 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3001 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3061 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3121 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3181 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3241 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3301 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3361 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3421 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     3481 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3541 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3601 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3661 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     3721 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3781 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3841 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3901 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3961 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4021 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4081 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4141 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4201 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4261 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg
     4321 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4381 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4441 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4501 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4561 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4621 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4681 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4741 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4801 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4861 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4921 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4981 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5041 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5101 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5161 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5221 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5281 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5341 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5401 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5461 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5521 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5581 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5641 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5701 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5761 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5821 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5881 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5941 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6001 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6061 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6121 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6181 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6241 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6301 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6361 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6421 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6481 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6541 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6601 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6661 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6721 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6781 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     6841 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     6901 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     6961 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     7021 ggccagatat acgcgtt
//
