LOCUS       sequence-203072-        4143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  entry clone for gateway cloning, cell cycle marker proliferative 
            cell nuclear antigen PCNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4143)
  AUTHORS   Icha J, Kunath C, Rocha-Martins M, Norden C
  TITLE     Independent modes of ganglion cell translocation ensure correct 
            lamination of the zebrafish retina.
  JOURNAL   J Cell Biol. 2016 Oct 24;215(2):259-275.
  PUBMED    27810916
REFERENCE   2  (bases 1 to 4143)
  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: "J Cell 
            Biol."; date: "2016-10-24"; volume: "215(2)"; pages: "259-275"
COMMENT     Sequence Label: pME GFP-PCNA
FEATURES             Location/Qualifiers
     source          1..4143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             671..691
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      704..713
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(710..712,713..715,716..1426)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(755..776)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1016..1035)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1363..1384
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2268..2367)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2384..2403)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2385..2403)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2408..2424)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2408..2424)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2526..2545)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2537..3346
                     /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     complement(2604..2623)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3493..4081
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3982..4001
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgg ccgaagaaga agcgcaaagt actggtaccg gtcgccacca tggtgagcaa
      721 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      781 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      841 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
      901 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
      961 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1021 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1081 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1141 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggc
     1201 caacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1261 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1321 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1381 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggcg aagggcaagg
     1441 gcaagggcaa gggccgggcc gcggctacgc gtatcgatcc atgttcgagg cgcgcctggt
     1501 ccagggctcc atcctcaaga aggtgttgga ggcactcaag gacctcatca acgaggcctg
     1561 ctgggatatt agctccagcg gtgtaaacct gcagagcatg gactcgtccc acgtctcttt
     1621 ggtgcagctc accctgcggt ctgagggctt cgacacctac cgctgcgacc gcaacctggc
     1681 catgggcgtg aacctcacca gtatgtccaa aatactaaaa tgcgccggca atgaagatat
     1741 cattacacta agggccgaag ataacgcgga taccttggcg ctagtatttg aagcaccaaa
     1801 ccaggagaaa gtttcagact atgaaatgaa gttgatggat ttagatgttg aacaacttgg
     1861 aattccagaa caggagtaca gctgtgtagt aaagatgcct tctggtgaat ttgcacgtat
     1921 atgccgagat ctcagccata ttggagatgc tgttgtaatt tcctgtgcaa aagacggagt
     1981 gaaattttct gcaagtggag aacttggaaa tggaaacatt aaattgtcac agacaagtaa
     2041 tgtcgataaa gaggaggaag ctgttaccat agagatgaat gaaccagttc aactaacttt
     2101 tgcactgagg tacctgaact tctttacaaa agccactcca ctctcttcaa cggtgacact
     2161 cagtatgtct gcagatgtac cccttgttgt agagtataaa attgcggata tgggacactt
     2221 aaaatactac ttggctccca agatcgagga tgaagaagga tcttaggacc cagctttctt
     2281 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2341 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2401 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2461 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2521 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2581 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2641 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2701 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2761 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2821 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     2881 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     2941 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     3001 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     3061 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3121 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3181 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3241 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3301 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3361 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3421 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3481 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3541 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3601 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3661 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3721 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3781 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3841 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3901 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3961 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4021 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4081 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4141 gtt
//
