LOCUS       Exported                6987 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    GFP-Nucleolin
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6987)
  AUTHORS   Takagi M, Absalon MJ, McLure KG, Kastan MB
  TITLE     Regulation of p53 translation and induction after DNA damage by 
            ribosomal protein L26 and nucleolin.
  JOURNAL   Cell. 2005 Oct 7. 123(1):49-63.
  PUBMED    16213212
REFERENCE   2  (bases 1 to 6987)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6987
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             620..1336
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             620..1336
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(665..686)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(926..945)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1273..1294
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1931..1942
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     protein_bind    1949..1960
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     CDS             3008..3019
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3523..3657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3782..3903
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3910..4365)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4047..4068)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4259..4278
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4392..4496
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4498..4855
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4521..4541)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4706..4841
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4768..4787
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4890..5684
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4944..4963)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5554..5573
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5872..5891)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5916..5963
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6292..6880
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6781..6800
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      661 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      721 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      781 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
      841 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      901 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
      961 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1021 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1081 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1141 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1201 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     1261 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1321 ggacgagctg tacaagtact cagatctcga gctcaagctt ggtaccatgg tgaagctcgc
     1381 gaaggcaggt aaaaatcaag gtgaccccaa gaaaatggct cctcctccaa aggaggtaga
     1441 agaagatagt gaagatgagg aaatgtcaga agatgaagaa gatgatagca gtggagaaga
     1501 ggtcgtcata cctcagaaga aaggcaagaa ggctgctgca acctcagcaa agaaggtggt
     1561 cgtttcccca acaaaaaagg ttgcagttgc cacaccagcc aagaaagcag ctgtcactcc
     1621 aggcaaaaag gcagcagcaa cacctgccaa gaagacagtt acaccagcca aagcagttac
     1681 cacacctggc aagaagggag ccacaccagg caaagcattg gtagcaactc ctggtaagaa
     1741 gggtgctgcc atcccagcca ggggggcaaa gaatggcaag aatgccgagg aggaagacag
     1801 tgatgaagag gaggatgatg acagtgagga ggatgaggag gatgacgagg acgaggatga
     1861 ggatgaagat gaaattgaac cagcagcgat gaaagcagca gctgctgccc ctgcctcaga
     1921 ggatgaggac gatgaggatg acgaagatga tgaggatgac gatgacgatg aggaagatga
     1981 ctctgaagaa gaagctatgg agactacacc agccaaagga aagaaagctg caaaagttgt
     2041 tcctgtgaaa gccaagaacg tggctgagga tgaagatgaa gaagaggatg atgaggacga
     2101 ggatgacgac gacgacgaag atgatgaaga tgatgatgat gaagatgatg aggaggagga
     2161 agaagaggag gaggaagagc ctgtcaaaga agcacctgga aaacgaaaga aggaaatggc
     2221 caaacagaaa gcagctcctg aagccaagaa acagaaagtg gaaggcacag aaccgactac
     2281 ggctttcaat ctctttgttg gaaacctaaa ctttaacaaa tctgctcctg aattaaaaac
     2341 tggtatcagc gatgtttttg ctaaaaatga tcttgctgtt gtggatgtca gaattggtat
     2401 gactaggaaa tttggttatg tggattttga atctgctgaa gacctggaga aagcgttgga
     2461 actcactggt ttgaaagtct ttggcaatga aattaaacta gagaaaccaa aaggaaaaga
     2521 cagtaagaaa gagcgagatg cgagaacact tttggctaaa aatctccctt acaaagtcac
     2581 tcaggatgaa ttgaaagaag tgtttgaaga tgctgcggag atcagattag tcagcaagga
     2641 tgggaaaagt aaagggattg cttatattga atttaagaca gaagctgatg cagagaaaac
     2701 ctttgaagaa aagcagggaa cagagatcga tgggcgatct atttccctgt actatactgg
     2761 agagaaaggt caaaatcaag actatagagg tggaaagaat agcacttgga gtggtgaatc
     2821 aaaaactctg gttttaagca acctctccta cagtgcaaca gaagaaactc ttcaggaagt
     2881 atttgagaaa gcaactttta tcaaagtacc ccagaaccaa aatggcaaat ctaaagggta
     2941 tgcatttata gagtttgctt cattcgaaga cgctaaagaa gctttaaatt cctgtaataa
     3001 aagggaaatt gagggcagag caatcaggct ggagttgcaa ggacccaggg gatcacctaa
     3061 tgccagaagc cagccatcca aaactctgtt tgtcaaaggc ctgtctgagg ataccactga
     3121 agagacatta aaggagtcat ttgacggctc cgttcgggct aggatagtta ctgaccggga
     3181 aactgggtcc tccaaagggt ttggttttgt agacttcaac agtgaggagg atgccaaagc
     3241 tgccaaggag gccatggaag acggtgaaat tgatggaaat aaagttacct tggactgggc
     3301 caaacctaag ggtgaaggtg gcttcggggg tcgtggtgga ggcagaggcg gctttggagg
     3361 acgaggtggt ggtagaggag gccgaggagg atttggtggc agaagccggg gaggctttgg
     3421 agggcgagga ggcttccgag gaggcagagg aggaggaggt gaccacaagc cacaaggaaa
     3481 gaagacgaag tttgaatagc tcgagcatgc atctagagga tcaacttgtt tattgcagct
     3541 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3601 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatccac
     3661 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     3721 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     3781 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3841 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3901 ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     3961 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     4021 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     4081 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     4141 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     4201 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     4261 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     4321 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     4381 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4441 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     4501 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4561 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4621 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4681 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4741 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4801 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     4861 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4921 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4981 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5041 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     5101 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5161 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5221 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5281 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5341 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5401 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5461 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5521 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5581 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5641 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5701 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5761 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5821 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     5881 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     5941 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     6001 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     6061 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     6121 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     6181 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6241 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6301 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6361 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6421 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6481 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6541 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6601 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6661 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6721 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6781 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6841 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6901 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6961 tgattctgtg gataaccgta ttaccgc
//
