LOCUS       sequence-431440-        4525 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (C' terminus) SV40NLS; Linker; p300.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4525)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 4525)
  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: "Cell 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: AA141
FEATURES             Location/Qualifiers
     source          1..4525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             444..464
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             513..2363
                     /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      2127..2136
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2435..2451)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2435..2451)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2448..2470)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2459..2475
                     /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(2483..2489,2490..2507,2508..2513))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2528..2549
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2666..2683)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2837..3425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2917..2936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3603..4412)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4326..4345
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4404..4423
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgagctcggt acctcgcgaa
      421 tacatctaga tgaagacggg tccccaaaga aaaagcggaa agtaagcgga agtgagaccc
      481 caggtacatc cgaatcagca acgcctgaaa gcattttcaa accagaagaa ctacgacagg
      541 cactgatgcc aactttggag gcactttacc gtcaggatcc agaatccctt ccctttcgtc
      601 aacctgtgga ccctcagctt ttaggaatcc ctgattactt tgatattgtg aagagcccca
      661 tggatctttc taccattaag aggaagttag acactggaca gtatcaggag ccctggcagt
      721 atgtcgatga tatttggctt atgttcaata atgcctggtt atataaccgg aaaacatcac
      781 gggtatacaa atactgctcc aagctctctg aggtctttga acaagaaatt gacccagtga
      841 tgcaaagcct tggatactgt tgtggcagaa agttggagtt ctctccacag acactgtgtt
      901 gctacggcaa acagttgtgc acaatacctc gtgatgccac ttattacagt taccagaaca
      961 ggtatcattt ctgtgagaag tgtttcaatg agatccaagg ggagagcgtt tctttggggg
     1021 atgacccttc ccagcctcaa actacaataa ataaagaaca attttccaag agaaaaaatg
     1081 acacactgga tcctgaactg tttgttgaat gtacagagtg cggaagaaag atgcatcaga
     1141 tctgtgtcct tcaccatgag atcatctggc ctgctggatt cgtctgtgat ggctgtttaa
     1201 agaaaagtgc acgaactagg aaagaaaata agttttctgc taaaaggttg ccatctacca
     1261 gacttggcac ctttctagag aatcgtgtga atgactttct gaggcgacag aatcaccctg
     1321 agtcaggaga ggtcactgtt agagtagttc atgcttctga caaaaccgtg gaagtaaaac
     1381 caggcatgaa agcaaggttt gtggacagtg gagagatggc agaatccttt ccataccgaa
     1441 ccaaagccct ctttgccttt gaagaaattg atggtgttga cctgtgcttc tttggcatgc
     1501 atgttcaaga gtatggctct gactgccctc cacccaacca gaggagagta tacatatctt
     1561 acctcgatag tgttcatttc ttccgtccta aatgcttgag gactgcagtc tatcatgaaa
     1621 tcctaattgg atatttagaa tatgtcaaga aattaggtta cacaacaggg catatttggg
     1681 catgtccacc aagtgaggga gatgattata tcttccattg ccatcctcct gaccagaaga
     1741 tacccaagcc caagcgactg caggaatggt acaaaaaaat gcttgacaag gctgtatcag
     1801 agcgtattgt ccatgactac aaggatattt ttaaacaagc tactgaagat agattaacaa
     1861 gtgcaaagga attgccttat ttcgagggtg atttctggcc caatgttctg gaagaaagca
     1921 ttaaggaact ggaacaggag gaagaagaga gaaaacgaga ggaaaacacc agcaatgaaa
     1981 gcacagatgt gaccaaggga gacagcaaaa atgctaaaaa gaagaataat aagaaaacca
     2041 gcaaaaataa gagcagcctg agtaggggca acaagaagaa acccgggatg cccaatgtat
     2101 ctaacgacct ctcacagaaa ctatatgcca ccatggagaa gcataaagag gtgttctttg
     2161 tgatccgcct cattgctggc cctgctgcca actccctgcc tcccattgtt gatcctgatc
     2221 ctctcatccc ctgcgatctg atggatggtc gggatgcgtt tctcacgctg gcaagggaca
     2281 agcacctgga gttctcttca ctccgaagag cccagtggtc caccatgtgc atgctggtgg
     2341 agctgcacac gcagagccag gacggaagca cgtcttcatc ggatcccggg cccgtcgact
     2401 gcagaggcct gcatgcaagc ttggtgtaat catggtcata gctgtttcct gtgtgaaatt
     2461 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     2521 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2581 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2641 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2701 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2761 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2821 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2881 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2941 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3001 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3061 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3121 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3181 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3241 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3301 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3361 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3421 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3481 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3541 aaaaatgaag ttttaaatca agcccaatct gaataatgtt acaaccaatt aaccaattct
     3601 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     3661 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     3721 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     3781 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     3841 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     3901 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     3961 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     4021 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     4081 tattcttcta atacctggaa tgctgttttt ccggggatcg cagtggtgag taaccatgca
     4141 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     4201 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     4261 aacaactctg gcgcatcggg cttcccatac aagcgataga ttgtcgcacc tgattgcccg
     4321 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     4381 ggcctcgacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     4441 gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga
     4501 ttttgagaca cgggccagag ctgca
//
