LOCUS       sequence-365219-        7047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PhCMV-driven histone acetyltransferase p300 
            core domain expression (PhCMV-p300core-pA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7047)
  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 7047)
  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: pVH19-Tier1-PhCMV-p300core
FEATURES             Location/Qualifiers
     source          1..7047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        113..492
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        493..696
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     647..667
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     741..760
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        741..759
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             806..2656
                     /codon_start=1
                     /gene="EP300"
                     /product="catalytic histone acetyltransferase core of the 
                     human E1A-associated protein p300 (Delvecchio et al., 
                     2013)"
                     /label=p300 core
                     /translation="IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDI
                     VKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQ
                     EIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQG
                     ESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAG
                     FVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHA
                     SDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPP
                     PNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDY
                     IFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFE
                     GDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS
                     RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDL
                     MDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQD"
     regulatory      2420..2429
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2711..2728)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2717..2941
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2987..3415
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3074..3093)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3284..3305
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3424..3444)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3429..3758
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3609..3744
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3671..3690
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3825..4619
                     /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(3879..3898)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4489..4508
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4793..4914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4830..4849)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4884..4903
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4963..4979)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4963..4979)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4976..4998)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4987..5003
                     /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(5011..5017,5018..5035,5036..5041))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5056..5077
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5194..5211)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5365..5953)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5445..5464)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6124..6915,6916..6984))
                     /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     6747..6766
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6985..7047,1..42))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
       61 gccacctgac gtcgacggat cgggagatct ccacgcgtgg taccctcgag ttgacattga
      121 ttattgagta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      181 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      241 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      301 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      361 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      421 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      481 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      541 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      601 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      661 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      721 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc tgttcgaagc
      781 ggaattcacc atgactagtc gcgccatttt caaaccagaa gaactacgac aggcactgat
      841 gccaactttg gaggcacttt accgtcagga tccagaatcc cttccctttc gtcaacctgt
      901 ggaccctcag cttttaggaa tccctgatta ctttgatatt gtgaagagcc ccatggatct
      961 ttctaccatt aagaggaagt tagacactgg acagtatcag gagccctggc agtatgtcga
     1021 tgatatttgg cttatgttca ataatgcctg gttatataac cggaaaacat cacgggtata
     1081 caaatactgc tccaagctct ctgaggtctt tgaacaagaa attgacccag tgatgcaaag
     1141 ccttggatac tgttgtggca gaaagttgga gttctctcca cagacactgt gttgctacgg
     1201 caaacagttg tgcacaatac ctcgtgatgc cacttattac agttaccaga acaggtatca
     1261 tttctgtgag aagtgtttca atgagatcca aggggagagc gtttctttgg gggatgaccc
     1321 ttcccagcct caaactacaa taaataaaga acaattttcc aagagaaaaa atgacacact
     1381 ggatcctgaa ctgtttgttg aatgtacaga gtgcggaaga aagatgcatc agatctgtgt
     1441 ccttcaccat gagatcatct ggcctgctgg attcgtctgt gatggctgtt taaagaaaag
     1501 tgcacgaact aggaaagaaa ataagttttc tgctaaaagg ttgccatcta ccagacttgg
     1561 cacctttcta gagaatcgtg tgaatgactt tctgaggcga cagaatcacc ctgagtcagg
     1621 agaggtcact gttagagtag ttcatgcttc tgacaaaacc gtggaagtaa aaccaggcat
     1681 gaaagcaagg tttgtggaca gtggagagat ggcagaatcc tttccatacc gaaccaaagc
     1741 cctctttgcc tttgaagaaa ttgatggtgt tgacctgtgc ttctttggca tgcatgttca
     1801 agagtatggc tctgactgcc ctccacccaa ccagaggaga gtatacatat cttacctcga
     1861 tagtgttcat ttcttccgtc ctaaatgctt gaggactgca gtctatcatg aaatcctaat
     1921 tggatattta gaatatgtca agaaattagg ttacacaaca gggcatattt gggcatgtcc
     1981 accaagtgag ggagatgatt atatcttcca ttgccatcct cctgaccaga agatacccaa
     2041 gcccaagcga ctgcaggaat ggtacaaaaa aatgcttgac aaggctgtat cagagcgtat
     2101 tgtccatgac tacaaggata tttttaaaca agctactgaa gatagattaa caagtgcaaa
     2161 ggaattgcct tatttcgagg gtgatttctg gcccaatgtt ctggaagaaa gcattaagga
     2221 actggaacag gaggaagaag agagaaaacg agaggaaaac accagcaatg aaagcacaga
     2281 tgtgaccaag ggagacagca aaaatgctaa aaagaagaat aataagaaaa ccagcaaaaa
     2341 taagagcagc ctgagtaggg gcaacaagaa gaaacccggg atgcccaatg tatctaacga
     2401 cctctcacag aaactatatg ccaccatgga gaagcataaa gaggtcttct ttgtgatccg
     2461 cctcattgct ggccctgctg ccaactccct gcctcccatt gttgatcctg atcctctcat
     2521 cccctgcgat ctgatggatg gtcgggatgc gtttctcacg ctggcaaggg acaagcacct
     2581 ggagttctct tcactccgaa gagcccagtg gtccaccatg tgcatgctgg tggagctgca
     2641 cacgcagagc caggacgcta gcggatccac cggtgtctag aaagctttga ggccggcctg
     2701 cagggatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2761 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2821 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2881 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2941 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3001 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3061 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3121 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3181 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3241 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3301 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3361 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3421 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3481 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3541 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3601 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3661 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3721 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3781 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3841 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3901 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3961 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4021 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4081 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4141 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4201 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4261 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4321 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4381 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4441 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4501 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4561 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4621 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4681 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4741 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4801 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4861 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4921 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4981 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5041 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5101 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5161 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5221 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5281 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5341 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5401 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5461 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5521 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5581 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5641 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5701 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5761 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5821 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5881 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5941 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6001 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6061 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6121 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6181 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6241 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6301 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6361 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6421 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6481 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6541 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6601 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6661 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6721 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6781 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6841 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6901 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6961 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7021 gggttattgt ctcatgagcg gatacat
//
