LOCUS       sequence-427399-        6426 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6426)
  AUTHORS   Yun C, Wang Y, Mukhopadhyay D, Backlund P, Kolli N, Yergey A, 
            Wilkinson KD, Dasso M
  TITLE     Nucleolar protein B23/nucleophosmin regulates the vertebrate SUMO 
            pathway through SENP3 and SENP5 proteases.
  JOURNAL   J Cell Biol. 2008 Nov 17;183(4):589-95.
  PUBMED    19015314
REFERENCE   2  (bases 1 to 6426)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol."; date: "2008-11-17"; volume: "183(4)"; pages: "589-95"
COMMENT     Sequence Label: GFP-SENP3
FEATURES             Location/Qualifiers
     source          1..6426
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /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(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3154..3275
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3191..3210)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3245..3264
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3282..3737)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3419..3440)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3631..3650
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3764..3868
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3870..4227
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3870..4061
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3893..3913)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4078..4213
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4262..5056
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4262..5056
                     /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(4316..4335)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4926..4945
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5244..5263)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5288..5335
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5664..6252
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6153..6172
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt actcagatct cgagatgaaa gagactatac aagggaccgg gtcctggggg
     1321 cctgagcctc ctggacccgg cataccccca gcttactcaa gtcccaggcg ggagcgtctt
     1381 cgttggcccc cacctcccaa accccgactc aagtcaggtg gagggtttgg gccagatcct
     1441 gggtcaggga ccacagtgcc agccagacgc ctccctgtcc cccgaccctc ttttgatgcc
     1501 tcagcaagtg aagaggagga agaagaggag gaggaggagg atgaagatga agaggaggaa
     1561 gtggcagctt ggaggctgcc cccaagatgg agtcagctgg gaacctccca gcggccccgc
     1621 ccttctagac ccactcatcg aaaaacctgc tcacagcgcc gccgccgagc catgagagcc
     1681 ttccggatgc tgctctactc aaaaagcacc tcgctgacat tccactggaa gctttggggg
     1741 cgccaccggg gccggcggcg gggcctcgca caccccaaga accatctttc accccagcaa
     1801 gggggtgcga cgccacaggt gccatccccc tgttgtcgtt ttgactcccc ccgggggcca
     1861 cctccacccc ggctgggtct gctaggcgct ctcatggctg aggatggggt gagagggtct
     1921 ccaccagtgc cctctgggcc ccccatggag gaagatggac tcaggtggac tccaaagtct
     1981 cctctggacc ctgactcggg cctcctttca tgtactctgc ccaacggttt tgggggacaa
     2041 tctgggccag aaggggagcg cagcttggca ccccctgatg ccagcatcct catcagcaat
     2101 gtgtgcagca tcggggacca tgtggcccag gagctttttc agggctcaga tttgggcatg
     2161 gcagaagagg cagagaggcc tggggagaaa gccggccagc acagccccct gcgagaggag
     2221 catgtgacct gcgtacagag catcttggac gaattccttc aaacgtatgg cagcctcata
     2281 cccctcagca ctgatgaggt agtagagaag ctggaggaca ttttccagca ggagttttcc
     2341 accccttcca ggaagggcct ggtgttgcag ctgatccagt cttaccagcg gatgccaggc
     2401 aatgccatgg tgaggggctt ccgagtggct tataagcggc acgtgctgac catggatgac
     2461 ttggggacct tgtatggaca gaactggctc aatgaccagg tgatgaacat gtatggagac
     2521 ctggtcatgg acacagtccc tgaaaaggtg catttcttca atagtttctt ctatgataaa
     2581 ctccgtacca agggttatga tggggtgaaa aggtggacca aaaacgtgga catcttcaat
     2641 aaggagctac tgctaatccc catccacctg gaggtgcatt ggtccctcat ctctgttgat
     2701 gtgaggcgac gcaccatcac ctattttgac tcgcagcgta ccctaaaccg ccgctgccct
     2761 aagcatattg ccaagtatct acaggcagag gcggtaaaga aagaccgact ggatttccac
     2821 cagggctgga aaggttactt caaaatgaat gtggccaggc agaataatga cagtgactgt
     2881 ggtgcttttg tgttgcagta ctgcaagcat ctggccctgt ctcagccatt cagcttcacc
     2941 cagcaggaca tgcccaaact tcgtcggcag atctacaagg agctgtgtca ctgcaaactc
     3001 actgtgtgag gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc
     3061 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     3121 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     3181 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3241 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     3301 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3361 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3421 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3481 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3541 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3601 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3661 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3721 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3781 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3841 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3901 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3961 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4021 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4081 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4141 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4201 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     4261 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4321 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4381 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4441 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4501 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4561 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4621 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4681 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4741 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4801 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4861 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4921 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4981 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5041 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5101 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5161 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5221 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5281 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     5341 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     5401 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     5461 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     5521 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5581 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5641 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5701 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5761 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5821 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5881 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5941 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6001 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6061 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6121 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6181 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6241 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6301 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6361 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     6421 gttccg
//
