LOCUS       sequence-365220-        5901 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PhCMV-driven Gal4-VP16 transactivator 
            (PhCMV-Gal4-VP16-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5901)
  AUTHORS   Haellman V, Strittmatter T, Bertschi A, Stucheli P, Fussenegger M
  TITLE     A versatile plasmid architecture for mammalian synthetic biology 
            (VAMSyB).
  JOURNAL   Metab Eng. 2021 Jul;66:41-50. doi: 10.1016/j.ymben.2021.04.003. Epub
            2021 Apr 18.
  PUBMED    33857582
REFERENCE   2  (bases 1 to 5901)
  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: "Metab 
            Eng."; date: "2021-07"; volume: "66:41-50. doi"; pages: " 
            10.1016/j.ymben.2021.04.003. Epub 2021 Apr 18"
COMMENT     Sequence Label: pTS2374-Tier1-PhCMV-Gal4_VP16
FEATURES             Location/Qualifiers
     source          1..5901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        32..411
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        412..615
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     566..586
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     660..679
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        660..678
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             731..1171
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             1190..1423
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(1484..1501)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1490..1714
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1760..2188
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1847..1866)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2057..2078
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2197..2217)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2202..2531
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2382..2517
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2444..2463
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2598..3392
                     /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(2652..2671)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3262..3281
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3566..3687
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3603..3622)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3657..3676
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3736..3752)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3736..3752)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3749..3771)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3760..3776
                     /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(3784..3790,3791..3808,3809..3814))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3829..3850
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3967..3984)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4138..4726)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4218..4237)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4897..5688,5689..5757))
                     /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     5520..5539
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5758..5862)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gggagatctc cacgcgtggt accctcgagt tgacattgat tattgagtag ttattaatag
       61 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      121 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      181 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      241 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      301 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      361 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      421 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      481 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      541 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      601 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
      661 aatacgactc actataggga gacccaagct gttcgaagcg gaattcacca tgactagtgg
      721 tggttctggt atgaagctac tgtcttctat cgaacaagca tgcgatattt gccgacttaa
      781 aaagctcaag tgctccaaag aaaaaccgaa gtgcgccaag tgtctgaaga acaactggga
      841 gtgtcgctac tctcccaaaa ccaaaaggtc tccgctgact agggcacatc tgacagaagt
      901 ggaatcaagg ctagaaagac tggaacagct atttctactg atttttcctc gagaagacct
      961 tgacatgatt ttgaaaatgg attctttaca ggatataaaa gcattgttaa caggattatt
     1021 tgtacaagat aatgtgaata aagatgccgt cacagataga ttggcttcag tggagactga
     1081 tatgcctcta acattgagac agcatagaat aagtgcgaca tcatcatcgg aagagagtag
     1141 taacaaaggt caaagacagt tgactgtatc ggctagtggt ggttctggtg cccccccgac
     1201 cgatgtcagc ctgggggacg agctccactt agacggcgag gacgtggcga tggcgcatgc
     1261 cgacgcgcta gacgatttcg atctggacat gttgggggac ggggattccc cgggtccggg
     1321 atttaccccc cacgactccg ccccctacgg cgctctggat atggccgact tcgagtttga
     1381 gcagatgttt accgatgccc ttggaattga cgagtacggt ggggctagtg ctagcggatc
     1441 caccggtgtc tagaaagctt tgaggccggc ctgcagggat cagcctcgac tgtgccttct
     1501 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     1561 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     1621 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     1681 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     1741 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1801 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1861 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1921 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     1981 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2041 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2101 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2161 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     2221 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2281 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2341 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2401 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2461 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2521 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     2581 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2641 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2701 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2761 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2821 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     2881 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     2941 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3001 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3061 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3121 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3181 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3241 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3301 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3361 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3421 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3481 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3541 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     3601 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3661 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     3721 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3781 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3841 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3901 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3961 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4021 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4081 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4141 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4201 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4261 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4321 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4381 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4441 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4501 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4561 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4621 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4681 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4741 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4801 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4861 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4921 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4981 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5041 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5101 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5161 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5221 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5281 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5341 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5401 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5461 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5521 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5581 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5641 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5701 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5761 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5821 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5881 gccacctgac gtcgacggat c
//
