LOCUS       sequence-334731-        5439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and mEGFP-linker sequence for N-terminus tagging of 
            human PCNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5439)
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5439)
  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: 
            "Unpublished"
COMMENT     Sequence Label: AICSDP-116:PCNA-mEGFP
FEATURES             Location/Qualifiers
     source          1..5439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..137
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     129..146
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     130..146
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1158..1167
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1164..1166,1167..1169,1170..1880)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1209..1230)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1470..1489)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1817..1838
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2969..2985)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2969..2985)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2982..3004)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2993..3009
                     /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(3017..3023,3024..3041,3042..3047))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3062..3083
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3200..3217)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3371..3959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3451..3470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4130..4921,4922..4990))
                     /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     4753..4772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4991..5095)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5163..5181
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5219..5241)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5341..5360
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
       61 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
      121 cacgacgttg taaaacgacg gccagtgaat tgacgcgtat tgggatttaa atgtgtacgc
      181 tctttgactc agctgtatta cttcaaggag ttgatatcac caaaattgcc taagtgctca
      241 aaggtgtttg tagttaaaca acaggagatt gataaattat gttatataca tgtgatgcta
      301 tgttttaaag aggtactgat atgataaaag atgtacgtgg cataaaatta aatgtacttt
      361 attaagtact tttccaagtg tttacggaat gagtgcattt ttgaaaaaaa aaaagtgtat
      421 tcgaactttt aaaaaagctt taaaagcttt atacaatgaa cgattgagtg attataagag
      481 ctggcggggg aatgttaaga ggatgatagg gagctaagtt taacagaaca attcacctct
      541 ttatcttgtg acacctacga gcgcatcaat tctgtaattg aaaaataaag tgcatatttg
      601 cagcagctgt actctcttca ggctgcaagg aggcttttcc tcccggtagg cttgatttgc
      661 atttcacttt cactttcgtg gctggaaact ttctacccac gtagtgggag gctgaggagc
      721 caccataaag ctggggcttg acgagccggg accgggaccc gatctccaca tatgcccgga
      781 cttgttctgc ggccgggttc aggagtcaaa gaggcgggga gacctgcgcg acgctgcccc
      841 gccctgcgcc cgcttcctcc aatgtatgct ctagggggcg ggcctcgcgg ggagcatgga
      901 cacgattggc cctaaagtct tccccgcaag gccgtgggct ggacagcgtg gtgacgtcgc
      961 aacgcggcgc agggtgagag cgcgcgcttg cggacgcggc ggcattaaac ggttgcaggc
     1021 gtagcagagt ggtcgttgtc tttctaggtc tcagccggtc gtcgcgacgt tcgcccgctc
     1081 gctctgaggc tcctgaagcc gaaaccagct agactttcct ccttcccgcc tgcctgtagc
     1141 ggcgttgttg ccactccgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1201 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1261 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1321 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1381 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1441 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1501 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1561 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1621 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1681 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1741 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     1801 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1861 gcatggacga gctgtacaag ggcgaaggac agggccaggg acagggtcca ggaagagggt
     1921 acgcctatag atctttcgag gcgcgcctcg tgcagggctc catcctcaag aaggtgttgg
     1981 aggcactcaa ggacctcatc aacgaggcct gctgggatat tagctccagc ggtgtaaacc
     2041 tgcagagcat ggactcgtcc cacgtctctt tggtgcagct caccctgcgg tctgagggct
     2101 tcgacaccta ccgctgcgac cgcaacctgg ccatgggcgt gaacctcacc aggtgagcct
     2161 cgcggccccg ggaagccggc cccggcccgc ctgcacctcc ggtgcttggc gggagcgctt
     2221 tcgagcctag ccctcattgg ctggcgtggg ccatccgcgc cttctcattg gcctgccacg
     2281 cagtggggtg gggcccagct gagcgcgcgg ttcggaaaag cccgcgctgg ctgctgcgcg
     2341 aacctgcttt ttcgcgccaa agtcacaaag cgggtggtgg cgggaaaatc aagggttttt
     2401 ccgcagtgcc aggaacactg ttccagggct ctttgctcac taaatcctgt tggccttgaa
     2461 tggacgcttt agctgtggct ttcttgtttc tgagacggtc tcggtgtgtt gcccgggctg
     2521 gtctccaact tctgggctca agcgatcctc ccggctcagt cgcggttgac tttaaatgct
     2581 ttataatgcc cttgcgagaa atgtggcagc ctgtcatcct acttagtggc aggagattgt
     2641 ttctatccag aagggacact gctggtggta ttttagtata aatactgcca gatgcgtccc
     2701 aaaacgtctg cattaataat ggcatcctcc agcagtccgc ttaccctcca ccagttctga
     2761 gacggcctga tgggtgagag tggtaacccc ttctaaccgc gttcgaaata cagcccttca
     2821 gcagacggcg ttgattttaa agcatgtgtc tcctgtcttc tagtatgtcc aaaatactaa
     2881 aatgcgccgg caatgaagat atcattacac taagggccga agataacgcg gataatccca
     2941 atggcgcgcc gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3001 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3061 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3121 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3181 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3241 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3301 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3361 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3421 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3481 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3541 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3601 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3661 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3721 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3781 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3841 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3901 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3961 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4021 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4081 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4141 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4201 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4261 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4321 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4381 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4441 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4501 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4561 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4621 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4681 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4741 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4801 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4861 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4921 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4981 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5041 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5101 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     5161 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     5221 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     5281 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     5341 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     5401 gcgtaaggag aaaataccgc atcaggcgcc attcgccat
//
